Journal of Cell Science ta. 00 euie l,21;Uzm ta. 01.I a been has It 2011). al., the et Uezumi question 2010; al., (Joe et lineage strongly Uezumi adipogenic 2010; the muscle al., to et cells skeletal satellite of mouse data diversion recent apparent of other However, studies 2010). al., et from Sarig 2007; al., et (Brack VI desmin ernlcl ye ne h prpit utr conditions culture appropriate the even under and vitro and types lineage osteogenic myogenic cell fibroblastic, the neuronal from adipogenic, diverted 2006; into be al., can transformed et 2013) Zammit al., 2001; et Beauchamp, Yin has and muscle skeletal It (Zammit adult incomplete cells 2009). are 2002; which stem al., cells, Smith, et satellite and that and Delmonico suggested Ravussin been 2007; obesity Wagatsuma, 1983; 2003; al., sarcopenia, Tyler, et myopathies, (Borkan hallmark regeneration al., a many is et muscle some Vettor skeletal of in in 2006; degeneration the fibres) fatty al., Indeed, in muscle 2009). et located (replacing (Schrauwen-Hinderling normal fascicles adipocytes in pathologies within uncommon of not or form amounts, muscle) lipid modest the (in intramyocellular in space physiological perimysial as or either Fat be pathological) either can al., 2006). which or be al., et fibres, muscle et can Vettor (within Coppi 2008; droplets muscle al., (De al., et in muscle et Shefer skeletal deposition 2002; Aguiari human al., 2005; Asakura et and al., 1997; Wada 2009) et 2001; al., al., Vertino et et 2004; (Grimaldi Csete 2001; mouse source Sambasivan al., the in muscle as et 2011; proposed accumulation been in al., also fat have 1961) et cells of these (Mauro, (Lepper but 2011), known cells al., et well satellite is of regeneration role essential The Introduction words: Key at even that demonstrated (CD56 and cells digestion, myogenic enzymatic of by predominantly muscle consisted skeletal cultures human these from isolation isolated after types hours cell 72 adherent the characterised We Summary 10.1242/jcs.132563 doi: 5610–5625 126, ß Science Cell of Journal 2013 September 9 Accepted Harridge* Campus, Guy’s R. House, D. Shepherd’s ( London, Stephen correspondence College and for King’s *Author Sciences, Lazarus Biomedical UK R. of 1UL, School Norman SE1 Sciences, Velloso, Physiological London P. Aerospace and Cristiana Human Rowlerson, of Centre M. Anthea Agley, C. differentiation Chibeza adipogenic myogenic undergo not readily but cells, fibroblasts, muscle skeletal Human 5610 irbat odfeetaeit dpcts ssonb oso nrclua E7 peuaino h dpgnctasrpinfactors transcription adipogenic the of upregulation the caused TE-7, treatments intracellular Both of could medium. loss populations adipocyte-inducing by cell or shown purified acids as the fatty adipocytes, if either PPAR into tested with we differentiate treatment factors, to to transcription response fibroblasts and in markers adipocytes lineage into content, differentiate lipid for staining immunofluorescent elsrigmto o h eaaino ygncclsfo irbat.Ti rcdr aeprte of purities gave procedure This fibroblasts. from cells myogenic of separation the for method cell-sorting hwddfeeta euaino PPAR of regulation differential showed ucefbolssaea es ioetpoeiosta a eana xrclua-arxpouigclso ifrnit into differentiate or cells extracellular-matrix-producing as remain can that progenitors bipotent least at adipocytes. are fibroblasts muscle 03 ulse yTeCmayo ilgssLtd Biologists of Company The by Published 2013. + PDGFR , Aauae l,20;Wd ta. 02 n also and 2002) al., et Wada 2001; al., et (Asakura c + n C/EBP and el.TeCD56 The cells. ) irbat,Aioeei,Se el,Seea uce aelt el,Transdifferentiation cells, Satellite muscle, Skeletal cells, Stem Adipogenesis, Fibroblasts, a + vimentin , a n dpino ii-ae dpct opooy ycnrs,moei el i o neg dpgnssand adipogenesis undergo not did cells myogenic contrast, By morphology. adipocyte lipid-laden a of adoption and , [email protected] + 2 fibronectin , rcinotie rmtefrtsr a ihyerce nTE-7 in enriched highly was sort first the from obtained fraction ) + .T vlaetebhvoro h eltpsotie,w piie obeimmuno-magnetic double a optimised we obtained, types cell the of behaviour the evaluate To ). c n C/EBP and a nrsos oteeaioei ramns u eut hwta ua skeletal human that show results Our treatments. adipogenic these to response in nvivo in in antccl-otn ehdbsdo h cell-surface the on immuno- based double a method optimised we cell-sorting this achieve To Catalioto magnetic 2008; 2009). to (AIM) al., al., et medium Rajashekhar et adipocyte-inducing (Watt muscle 2008; an Moelling, (FAs) second, and human (Fritzius and acids 2012) a fatty al., first, from et stimuli; cue, different nutrient fibroblasts two physiologically-relevant using adipocytes and into differentiate cells myogenic 1984; 2008). al., al., et et Goodpaster (Haynes 2003; al., muscle et skeletal Stewart human including tissues, validated al., carefully situ et been Pfisterer in has 2009; antigen al., TE-7 (Stewart et the Pilling Furthermore, types 2008; 2011). al., cell et from Pollina other 2003; them al., and et distinguish macrophages and chondrocytes, endothelial fibroblasts cells, fibrocytes, monocyte-derived identify blood to peripheral al., human that used et protein tissue Mathew be connective 2008; fibroblast-specific can al., an a et is but TE-7 (Goodpaster specific 2011). cells problematic, identifying these on be for concentrated has markers can work of culture body increasing in between 2011). fibroblasts relationship al., et (Murphy the Identifying unclear is However, fibroblasts stromal al., 2010). local were and et al., FAPs (Joe muscle et adipocytes Uezumi skeletal or fibroblasts 2010; mouse into differentiate from to reported isolated 2011). 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(age the old around an staining from TE-7 muscle strong skeletal also of cryosection and wh endomysi Typical (asterisk), staining) the perimysium (E) (Oil-Red-O throughout the C. deposition dispersed throughout in fat is present enlarged intramuscular also TE-7 is gross is that that of reactivity indicates field area TE-7 the staining (D) large shows laminin lamina. a box and basal showing white TE-7 laminin-positive the the of staining; of Comparison laminin surface (C) have outer B). the in with (enlarged merges lamina and basal the under position akye l,20)frterpouil uiiainand human from purification populations reproducible cell the major the CD56 for 1987; muscle. of al., 2009) et culture (Cashman al., subsequent cells et myogenic on Mackey CD56 of expression i.1 muooaiaino D6adT- ncyscin fhmnseea muscle. skeletal human of cryosections on TE-7 and CD56 of Immunolocalisation 1. Fig. cuuain hne ncl opooy n xrsinof expression and morphology, cell in changes accumulation, expression. w ihyaioei u-ouain nseea muscle. skeletal in sub-populations adipogenic for highly markers as two indicated been recently have these because factor antigens, (PDGFR growth (Lecourt platelet-derived terminal and CD15 alpha 2010) the al., receptor expressing et the Pisani cells 2010; of al., et trigger presence (Shao the our tested for adipocytes also to We cultures 2006). mature Spiegelman, and Rosen known into 1997; Lazar, (C/EBP and preadipocytes are of alpha differentiation factors nuclear protein transcription the (PPAR binding gamma measured peroxisome enhancer receptor adipogenesis, we of activator regulators proliferator addition, master the of In to expression exposure stimuli. following factors, adipogenic transcription and markers lineage dpgncdfeetainwsdtrie ylipid by determined was differentiation Adipogenic m D;100 (D); m 2 el eecnimda irbat yterTE-7 their by fibroblasts as confirmed were cells m (E). m a Uzm ta. 00 elsurface cell 2010) al., et (Uezumi ) a eas these because ) c n CCAAT/ and ) ucefo on n nedrypro Fg 1). 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(Fig. oeta,mkn hmtems ieysuc fadipocytes conditions. of of adipogenic Results range source a inherent in likely to muscle skeletal most an the in resistance the accumulate have unlike that dish them considerable that making culture fibroblasts showed potential, indicate the the data which to adipogenesis, Our attach cells, isolation). they myogenic post after fibroblast other hours soon of (72 battery markers a and TE-7 lineage express highly muscle. cells skeletal human These in population cell adherent non-myogenic u eut lal hwta irbat r h otabundant most the are fibroblasts that show clearly results Our rndfeetaino ucefbolss5611 fibroblasts muscle of Transdifferentiation + oae eet h aa aiaecsn auefibres mature encasing lamina basal the beneath located ) oaiaino ygnc(D6 n connective and (CD56) myogenic of localisation A aelt el r dniidb hi CD56 their by identified are cells Satellite (A) nsitu in oaiaino D6( myogenic (a CD56 of localisation + tts(ros and (arrows) status c osnot does ich m m ividual um, Journal of Cell Science hn5 o-ygnccls w asstruhaMACS a through passes two CD56 cells, for selecting column sorting) non-myogenic cell (magnetic-activated 5% than al., et muscle 2012). usual (Bonavaud hindlimb Yablonka-Reuveni, (C57BL/6) the humans mouse and within and (Danoviz for isolation, 2007) is al., authors et yield post Zheng other cell 1997; of week by This mass reported 2D). the 1 (Fig. and with range correlation At sample good tissue TE-7 cells. the a the showed confirming these yields expressed cell MyoD, of adherent or never lineage desmin divergent expressed cells never Myogenic fibroblasts 2A–C). (Fig. n oewt ula yD n irbat(TE-7 fibroblast and MyoD) nuclear with some and ula xrsino i7 nclso ohmoei (desmin myogenic both of cells by in Ki67) (indicated culture of proliferation in expression showed days nuclear condition) 7 after ‘before-sort’ cells (the muscle-derived of Immunostaining cells fibroblasts myogenic and of purification immuno-magnetic Double Fig. material (supplementary 2006) PDGFR Murphy Zeisberg, fibronectin S3A,C). and and TE-7 2008; S2E) 2008) Fig. (Kalluri S2A,B), PDGFR al., material al., (supplementary et S2D), 2010) Fig. (Li Leask, et Fig. vimentin material material S2C), (Zou (supplementary (supplementary Fig. high material by VI 2011), their (supplementary evaluated al., collagen by was et confirmed of markers were lineage expression of Fibroblasts expression cell immunostaining. of hours abundance the relative isolated, 72 the after quantify types, were To early fixed attached. cells had and during they coverslips once present muscle-derived collagen-coated are human onto types plated culture, cell which of establish stages to order cultures muscle-derived In human of characterisation Early staining muscle. CD15 human No of 1E). sections these (Fig. on tissue detectable connective was staining the TE-7 in deposition, strong lipid intramuscular was and fibrosis areas included both which of subject, 1D). elderly the (Fig. was from absent muscle TE-7 anti- was in Similarly, perimysium, staining overlapped laminin in whereas However, partially expressed, highly 1C). also (Fig. and staining laminin fibres muscle surrounding 5612 el cone o ital l fteahrn muscle-derived adherent S3D). the myogenic Fig. of material and Fig. all (supplementary fibroblasts virtually population S2C; dissociation, for post Fig. accounted hours cells material 72 absent (supplementary At completely S3D). cells was and the myogenic was fibroblasts TE-7 from for used, markers Fig. discriminating material fibroblast putative most (supplementary S2D; the also fibronectin Of Fig. S3B). expressed cells Fig. many material 2010) Myogenic and al., (supplementary et S3D) S2B–E). Boldrin vimentin 1992; Fig. intracellular al., expressed the et material and Ven der (supplementary CD56 (van marker myogenic desmin surface All marker S3D). the Fig. same expressed material connective cells the (supplementary cells the from these capacity (ECM)-producing confirming of matrix coverslips extracellular matched, and independent lineage closely tissue on were TE-7 VI for population Counts membrane. collagen cell the and on pattern staining punctate ihicdneo i7epeso esse otsrigi both in post-sorting a persisted 2G–J, expression Fig. in Ki67 typical seen of be a can incidence of As high 2F–J. fractions Fig. sorted in and shown are sort’ experiment ‘before the sorting of the cell composition The of numbers. cell each initial variable for with yields biopsies seven cell from steps the shows 2E Fig. required. were orlal banhmnmoei elppltoswt less with populations cell myogenic human obtain reliably To ora fCl cec 2 (24) 126 Science Cell of Journal a + el eeqatfe ae na intense an on based quantified were cells a Bne,2004; (Bonner, + phenotypes ) + cells + , CD56 odto.Aayi fmxdppltoso el demonstrated cells CD56 of all populations nearly that mixed of Analysis condition. eyfwlpddolt Fg A o o,B,weesteewas there whereas B), CD56 the row, in top staining 3A, Oil-Red-O greater (Fig. significantly droplets lipid few very otat CD56 contrast, fe as h rae rlfrto aeo irbat nthis in 95 fibroblasts a of rate produced proliferation had greater fraction the days, 2 After ot1CD56 sort-1 ifrn epne eecnevdi ie utrsobtained cultures These D,F). mixed CD56 row, in sort-2 second the conserved from 3A, were (Fig. responses time different this over droplets lipid xrsinwsfudi h CD56 the CD15 in of found evidence No was co- S5C). these expression Fig. of always identity material was myogenic (supplementary the cells CD15 confirming always CD56 S5A,B). with was expressed Fig. material and (supplementary rare extremely Seate l,20)admuemsl Bsaosie l,2008). al., et (Bosnakovski muscle muscle mouse human and both 2003) for al., data et (Stewart published matches also population sort before- the in respectively) non-myogenic, and (myogenic types cell hra hsmre a lotudtcal nCD56 on undetectable almost was marker this whereas hleg,clue eeepsdt hsooia oeand dose nutrient albumin. physiological serum relevant to a complexed CD56 of FAs unsaturated to physiologically to response saturated exposed of adipogenic ratio a were the cultures assess to challenge, To cells C). row, muscle-derived third 3A, (Fig. el eoie nitiaeTE-7 intricate an deposited cells (CD56 myogenic being cells (91 remaining population fibroblast etse hte ot2CD56 sort-2 whether tested acid We fatty to response in treatment fibroblasts and in cells upregulated myogenic highly are factors transcription Adipogenic CD56 condition, control untreated the treatment In acid in fatty stores to lipid response intracellular expand vastly Fibroblasts CD56 sort-2 PDGFR and In from CD15 subsets by identified cell muscle stem human adipogenic putative of Presence ancl ouain (86 populations cell main flpd(i.3,bto o) uhta hi yols was cytoplasm their that such row), Oil-Red-O with bottom filled completely 3A, almost (Fig. lipid of odtos(i.2) nhgl ofun odtos CD56 conditions, confluent highly In same the 2N). under (Fig. differentiation myogenic conditions for capacity no showed rtasern,CD56 insulin (without transferrin), medium or differentiation myogenic serum-free In splmnaymtra i.S5D). Fig. material (supplementary 2hus h CD56 the hours, 72 kltlmsl,wihcncmlt emnlmyogenic terminal complete CD56 sort-1 human can the from contrast, which cells In differentiation. myogenic muscle, identifies skeletal expression confirming CD56 2M), (Fig. that MHC expressed strongly which purity myotubes, high very in resulted sorting double (97 The shown). not ( splmnaymtra i.S) hc sadefining a is which S4), Fig. material (supplementary hrceitco irbat Sbtlie l,2001). al., et (Sabatelli fibroblasts of characteristic n 5 h ot1CD56 sort-1 The 6 ujcs;ti uiyicesdwt iei utr (data culture in time with increased purity this subjects); 4 1%, 2 + /TE-7 el n CD56 and cells n 5 2 + ipis ftesr- CD56 sort-2 the of biopsies) 5 /TE-7 cells, + + /desmin utrs xrsino h D5atgnwas antigen CD15 the of expression cultures, 2 + 2 n 2 /TE-7 irbat xrse h atradipogenic master the expressed fibroblasts 2 5 + el hwdsrn tiigfrPDGFR for staining strong showed cells ujcs.Tepooto fteemajor these of proportion The subjects). 5 2 rcin(i.3G,H). 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Fig. e etpg o legend. for page next See rndfeetaino ucefbolss5613 fibroblasts muscle of Transdifferentiation Journal of Cell Science xrse ytesm ecnaeo el nclue M ot2purified Sort-2 (M) culture. in cells of percentage CD56 same the by expressed hntpsadyed band G el eoesrig(ie composition (mixed cell sorting the before desmin showing Cells of mg, (G) cell- obtained. 291.8 typical yields of one and biopsy of phenotypes a successive Results from seven (F) starting cell to biopsy). experiment Total applied each purification (E) process for sample. 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Lane, medium’ insulin and ‘adipocyte nutrition Tang PPAR 1989; ‘adipocyte al., and et al., an an 1976) (Hauner et days Ho, also 3 from Student and insulin for with inhibitor, 2006) 1978; (Russell 1974), induction IBMX) al., cAMP-phosphodiesterase as treatment et Meuth, adipocyte (a known (Rubin and 1-methyl-3-isobutylxanthine dexamethasone for initial (Green 1975), 1980), protocol Kehinde, serum an and standard containing (Green involves a medium’ differentiation to which exposed preadipocytes were Cells medium induction Adipogenic CD56 the from Cells to plates. treatment on 24-well FA densities in equivalent coverslips at or glass plates C8919) at 96-well Sigma, plated into mg/ml; were well process (0.3 per sorting cells collagen-coated the 2000 of of stage density each a from suspensions Cell population. eete olce yfuhn h ounwt ASbfe fe twas it after buffer MACS with CD56 ‘sort-1 column the releasing the field, flushing magnetic the by from removed collected then were rdcn o-eand‘ot2CD56 ‘sort-2 non-retained a producing Dk KLd abigsie K.Sandcyscin eemutdi Dako (Dako). in medium DAPI mounted gold’ were with ‘anti-fade cryosections gold’ Dako Stained ‘anti-fade UK). in Cambridgeshire, mounting Ltd, before UK incubation dye (Dako labelled minute antibody (Sigma) 10 fluorescently 33342 and primary Hoechst species-specific S3) in Table Unlabelled with material (supplementary incubation antibodies. antibodies by secondary primary followed with was binding incubated and blocked iiASsprto ntadMC utsad(itniBoeh.Non- the were Biotech). these – (Miltenyi collected were multistand column the CD56 MACS through ‘sort-1 straight and passing cell cells 130- unit retained The Biotech, 500 Miltenyi separation buffer. 130- with column; Biotech, MiniMACS cell MACS equilibrated (Miltenyi (large of previously column filter the ml pre-separation 042-202) into a l dripped through and in passed 041-407) resuspended then buffer, was finally MACS solution in and resuspended again were cells centrifuged 657 pelleted The at beads. centrifuged antibody-conjugated and buffer MACS ot1CD56 sort-1 rlfrto eim hsrsle nmlrrto f13131oleic 1.3:1.3:1 of ratios culture 2012). molar cell al., for et recommended (Watt in range values the serum resulted within mimicking studies is which This acid:BSA, acid:palmitic medium. 300 and proliferation acid oleic eete eesdfo h ouna ecie bv ogv h ot2CD56 sort-2 the give to above described as column the from released then were m hcns.Tsu etoswr u n tie sbelow. as stained and cut were sections Tissue thickness. m ´ rez-Zacarı + 2 ouainwste asdtruhafeheulbae column, equilibrated fresh a through passed then was population el’ antclyrtie el aeldb h niC5 beads anti-CD56 the by labelled cells retained Magnetically cells’. ´ se l,19;Komne l,20) fe l te tiigwas staining other all after 2001), al., et Koopman 1992; al., et as + m n CD56 and amtcai opee oBAa 5m/lin mg/ml 15 at BSA to complexed acid palmitic M 6 ˚ ,20 .Atricbto,tecl ouinwsdltdwith diluted was solution cell the incubation, After C. 6 2 ,40 rcin eeepsdfr7 or o300 to hours 72 for exposed were fractions 6 2 g betvswt .5 .5 .5numerical 0.95 0.75, 0.25, with objectives rcin n eandCD56 retained and fraction’ o iue ocletclsbudto bound cells collect to minutes 6 for m fMC ufradhl na in held and buffer MACS of l 3 00% o 0mnts then minutes, 10 for (0.01%) c gnss(eCpie al., et Coppi (De agonists + elfato.This fraction. cell ’ + el,which cells, m M + a nacrac ihsgetdgieie o iia mgs(ose and used were (Rossner images images monochrome bit digital 16 for or analysis. 8 quantitative guidelines formatting unadjusted for raw, post-acquisition suggested Only All unsharp with 2004). 2). an Yamada, accordance radius: and slightly in 150–200, the altered the (amount: was was In applied in balance 2009). was colour (Waters, mask figures, background fluorescence and field for Local background potential used modifications. non-specific for exposures micrographs account in some to variation with measurements identical field 2012) from to al., subtracted described at et were previously same (Agley values as intensity the al. photographed performed et with was Agley stained analysis were by Image and session. culture, interest, microscopy in same time of same the label for with correspond maintained to experiment antibody. micrographs secondary to the tones of colour wavelength blue emission or the green red, pseudo assign etakL ajrmadC ob o ehia assistance. technical for Hobbs C. method. current and our Marjoram based L. we thank which We upon us, with procedure sorting oeta w rus infcnewsasse ytowymxdmodel mixed of two-way value alpha by an cases significant. and all assessed experiments statistically of In for adjustment. analysis was given Bonferroni are For post-hoc significance numbers with groups. (ANOVA) groups, variance between of two analysis differences compare than to more used were distributed) experiments independent biopsy. representative three of a from fields from 15 sampled least derived condition) at basis, cells from of and on nuclei cell-by-cell population type performed all large on cell a a made (per in were on view expression measurements cases, factor quantified all transcription In of objectively cells. range full be the could revealing grey-scale) averaged oass hne nclua ii otn nteCD56 the in content lipid cellular in changes assess of content To feature lipid cellular ‘Count’ of the Measurement using Inc.). wells Systems six (Adobe to replicate software at five after in from Extended captured its well and CS5 view were per in Photoshop plates of counted fields or were 96-well fields non-overlapping cells figure in 1000 non-overlapping of the least plated images At on all were exposures. six either for standardised fractions to marked condition sorted five is per the immunostaining, it analysed from differed, and Cells number plated legend. the were if wells experiments; replicate three expression Normally, marker for counting Cell upeetr aeilaalbeoln at http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.132563/-/DC1 online available material Supplementary acknowledged. also is London; Trust College Spurrell King’s from the studentship from the PhD support a and by funded designed was read C.A. authors biopsies, the Funding All writing muscle manuscript. and manuscript. the the final experiments the wrote approved N.L. took the and and C.V. designing experiments, S.H. A.R., manuscript. in manuscript. work, the involved wrote experimental and all were data undertook the experiments, analysed the designed C.A. contributions Author Be to grateful are We Acknowledgements Unpaired Statistics to corresponding layer the to transferred field then a was in PPAR nuclei for selection all Hoechst staining select live blue antibody to This for applied in and view. mask factors created selection of was transcription nuclei range adipogenic stained colour (Sigma) and representative 33342 myogenic a of nuclei, expression individual the expression measure factor To transcription nuclear of Quantification ple qal oalfed fve n estvl iciiae vnvr small very even discriminated sensitively and view of fields all to equally applied ofre h ygnco irbatiett fteclue htwr analysed. staining were TE-7 that or cultures Desmin the of cell. to identity per normalised fibroblast content and or measured lipid myogenic was average the view give confirmed of The to field reproducible. each number exactly in nuclear were staining that Oil-Red-O measurements of unbiased area give to droplets lipid ouain,aseii oorrneslcinms Alye l,21)was 2012) al., et (Agley Oil-Red-O for mask threshold selection a range select colour to created specific a populations, oesr titcmaaiiy rae n oto el rmtesame the from cells control and treated comparability, strict ensure To rndfeetaino ucefbolss5623 fibroblasts muscle of Transdifferentiation t tsso anWinyUtss(hndt eenon-normally were data (when U-tests Mann–Whitney or -tests ´ ne ´ c it hzu o hrn e antccell- magnetic her sharing for Chazaud dicte rC/EBP or a ota h loecn inl(a non- (raw signal fluorescent the that so , + ii rpes hsms ol be could mask This droplets. lipid P , .5wsacpe as accepted was 0.05 + n CD56 and 2 cell n Journal of Cell Science aha,N . oal,J,Wlmn .L n ae,J R. J. Sanes, and L. R. Wollman, J., Covault, R., N. Cashman, rc,A . oby .J,Ry . e,M,Ko .J,Kle,C n ad,T A. T. Rando, and C. Keller, J., C. Kuo, M., Lee, S., Roy, J., M. Conboy, S., A. Brack, onkvk,D,X,Z,L,W,Te,S,Cevr . elner,R .R and R. C. R. Perlingeiro, O., Cleaver, S., Thet, W., Li, Z., Xu, K. C. D., Silbert, Bosnakovski, and H. A. Robbins, G., S. Gerzof, E., D. Hults, A., G. Borkan, onr .C. J. Bonner, G. Bo Barlovatz-Meimon, and K. R. Gherardi, P., Thibert, S., E. Bonavaud, J. Morgan, and F. Muntoni, L., Boldrin, S. M. Rodeheffer, and R. Berry, M. Bergstro Rudnicki, and M. Komaki, A., Asakura, A. P. Grimaldi, and A. N. Abumrad, G., Ailhaud, F., Bonino, E.-Z., Amri, re,H n et,M. Meuth, and H. Green, gir,P,Lo . aa,B,Vnin,V,Rmsi . inh,K,Fazn C., Franzin, K., Bianchi, A., Rimessi, V., Vindigni, B., Zavan, S., Leo, P., Aguiari, re,H n eid,O. Kehinde, and H. Green, ge,C . els,C . aau,N .adHrig,S .R. D. S. Harridge, and R. N. Lazarus, P., F.-J., C. Velloso, Authier, C., C. S., Agley, Valable, G., Pallafacchina, F., Grand, Le R., Abou-Khalil, References 5624 odatr . ees-ilr . aed .R,Ase,S . Randolph- C., S. Aisner, R., M. Hameed, A., Legesse-Miller, E. T., D. Goodpaster, Kelley, and L. F. Thaete, H., B. Goodpaster, K. Moelling, and T. Fritzius, L., Perani, A., Innocenzi, E., Azzoni, D., M., Covarello, G., Piccoli, Maroli, C., A., Centobene, Dellavalle, L., Boldrin, A., Scarda, G., Milan, P., Coppi, De Z. Yablonka-Reuveni, and M. Danoviz, S. Giuliani, and A. C. Maggi, R.-M., Catalioto, emnc,M . ars .B,Vse,M,Pr,S . ory .B,Velasquez- B., M. Conroy, W., S. Park, M., Visser, B., T. Harris, J., M. Delmonico, Wold, and C. J. Doyle, S., Korsnes, Y., Wei, N., Slawny, J., Walikonis, M., Csete, nean .G. C. ¨nnemann, deinmlcl nnra,dnrae,admoahchmnmuscle. human myopathic and denervated, normal, in molecule adhesion 20) nrae n inln uigaigatr ucese elft and fate cell stem muscle alters aging during signaling fibrosis. increases Wnt Increased (2007). reporter. M. Kyba, tomography. computed by revealed 673-677. composition body in changes Age yoieGot atrRev. Factor Growth Cytokine and biopsy, muscle of matrix. site donors, of sex and and processing. proliferation before age yield, delay cell to of according variations rates culture: differentiation cell satellite muscle human Primary same? the really cells vivo. in lineage and patients kidney subjects, diarrhoea. normal chronic on study with A patients specimens. biopsy needle on analysis in m acids exhibit that fatty cells of differentiation. stem effects multipotential transcriptional receptors. mediates proliferator-activated that peroxisome Chem. to protein Homology a preadipocytes. of Cloning 1231. ifrnito nculture. in differentiation ifrnito fmsl-eie tmcells. stem al. muscle-derived et of R. Vettor, differentiation M., Rossato, R., Cortivo, ifrnito nclue I atr fetn h dps conversion. adipose the affecting Factors II. culture. in differentiation ellrcnttet:apiaint h esrmn fhmnmsl el in cells muscle human of labeled measurement fluorescently of the quantitation to and culture. application selection constituents: precise the cellular for method analysis A. Sotiropoulos, F., Chretien, self-renewal. S., cell Germain, satellite K., R. al. Gherardi, et A., M. Rudnicki, dniyn irbat nfrai-ie,prfi-meddtissue. paraffin-embedded A. formalin-fixed, Cytochem. H. in diabetes Coller, fibroblasts 2 identifying and type J. in and Habecker, obesity in resistance insulin with mellitus. associated is distribution generate and fibres muscle into differentiate al. muscle et cells. S. skeletal satellite Tajbakhsh, postnatal S., in Brunelli, resident R., Sambasivan, S., al. Antonini, et cells. satellite P. muscle Gamba, human C., of 1962-1973. potential Pagano, adipogenic C., the modifies Pilon, M., Pozzobon, In system. culture primary a in Press. analysis Humana Protocols and and isolation Methods for Myogenesis: methods and background 21 iyr . odeu . aii .M,Nvt,M,Nwa,A .e al et B. adipogenesis. promotes A. protein Newman, M., Nevitt, infiltration. M., tissue T. adipose Manini, Body and R., Aging, Health, Boudreau, P., Mieyer, culture. in adipogenesis and B. program. differentiation at the preadipocytes of white human stage subcutaneous early of an conversion adipose prevent inhibitors 481-489. , 20) xgnmdae euaino kltlmsl aelt elproliferation cell satellite muscle skeletal of regulation Oxygen-mediated (2001). m J. ¨m, 20) uorn n aarn nipitn1Te2sgaigpooe muscle promotes signaling 1/Tie-2 angiopoietin paracrine and Autocrine (2009). 270 a.Rv Neurol. Rev. Nat. .Hsohm Cytochem. Histochem. J. tmCells Stem m .Ci.Nutr. Clin. J. Am. ora fCl cec 2 (24) 126 Science Cell of Journal 2367-2371. , 56 20) rsetv slto fseea ucese el ihaPax7 a with cells stem muscle skeletal of isolation Prospective (2008). 16) uceeetoye nmn eemndb eto activation neutron by Determined man. in electrolytes Muscle (1962). 347-358. , 20) euaino DFadisrcposi irtcdiseases. fibrotic in receptors its and PDGF of Regulation (2004). a.Commun Nat. Differentiation a.Cl Biol. Cell Nat. Science 21) h olgnV-eae yptis ucemesits meets muscle myopathies: VI-related collagen The (2011). 26 .Hsohm Cytochem. Histochem. J. 3194-3204. , il Cell Biol. Cell 7 elSe Cell Stem Cell 20) ogtdnlsuyo ucesrnt,qaiy and quality, strength, muscle of study Longitudinal (2009). m .Ci.Nutr. Clin. J. Am. 317 379-390. , 71 17) netbihdpeaioecl ieadits and line cell pre-adipose established An (1974). .Cl.Physiol. Cell. J. 2 20) k-adFx1itrcigWD-repeat-FYVE Foxo1-interacting and Akt- (2008). 17) netbihdpedps elln n its and line cell preadipose established An (1975). 499. , 15 807-810. , 885-892. , 3 68 MOJ. EMBO 15 127-133. , cn.J ln a.Invest. Lab. Clin. J. Scand. 255-273. , 245-253. , 21) hrceiaino h dpct cellular adipocyte the of Characterization (2013). 60 302-308. , 89 428-438. , 20) nimnhsohmclmto for method immunohistochemical An (2008). e.J .DMro,p.2-2 e ok NY: York, New 21-52. pp. DiMario), X. J. (ed. 233-240. , 5 27 298-309. , 21) kltlmsl aelt cells: satellite muscle Skeletal (2012). 20) ihguoeidcsadipogenic induces glucose High (2008). 1399-1410. , oei,otoei,adadipogenic and osteogenic, yogenic, 21) r ua n os satellite mouse and human Are (2010). rc al cd c.USA Sci. Acad. Natl. Proc. 90 189 x.Cl Res. Cell Exp. 1579-1585. , 58 20) hmclydsic HDAC distinct Chemically (2009). 20) ucestlieclsare cells satellite Muscle (2001). 189-196. , 941-955. , 20) hg dps tissue adipose Thigh (2000). 4Suppl 14 315 20) Rosiglitazone (2006). 3267-3280. , 18) erlcell Neural (1987). 21) nimage An (2012). 21) Pericytes (2011). Diabetologia .Gerontol. J. ,1-110. ., .Histochem. J. Cell n.Neurol. Ann. 105 5 .Biol. J. 19-27. , 1226- , (1983). (1997). (1995). 38 49 .; , , rgie .M,Sa,C .adSl .S. H. Sul, and M. C. Smas, M., F. Gregoire, lo,L .adSrao P. Soriano, and E. L. Olson, E. P. Scherer, and R. A. Nawrocki, A. Mauro, alr,R n eseg M. Zeisberg, A. and M. Rudnicki, R. J., Kalluri, Wang, L., So, F., Grand, Le A., Natarajan, L., Yi, B., W. A. Joe, epr . atig,T .adFn C.-M. Fan, and A. T. Partridge, C., Lepper, A. Leask, K. M. I. Hesselink, Karagiannides, and G. and Schaart, J. R., Han, Koopman, T., Pirtskhalava, T., Tchkonia, L., J. Kirkland, ans .F,Sere .M,Lbc,D .adHnly .L. L. Hensley, and F. D. Lobach, M., R. Scearce, F., B. Haynes, Z. E. Amri, and D. Gaillard, H., Inadera, L., Teboul, A., P. Grimaldi, upy .M,Lwo,J . ahw .J,Hthsn .A n adn G. Kardon, and A. D. Hutcheson, J., S. Mathew, A., J. Lawson, S. E. M., F. M. Walsh, J. and Murphy, G. Morgan, J. and Dickson, V., F. Kirkness, J., Muntoni, Thompson, W., E., S. S. Moore, Xu, F., C. Adkin, J., Meng, A., J. Lawson, M., M. Murphy, J., A. Merrell, M., J. Hansen, J., S. Lastayo, Mathew, and G. Morrell, B., K. Foreman, E., L. Holm, Dibble, J., O., Addison, Bojsen-Moller, L., R. J., Marcus, Henriksson, N., Charifi, R. M., K. Kjaer, Wagner, L., and A. D. Mackey, H. Kollias, B., Z. Li, en,H . ak .W,Km . ak .K n on D. Yoon, and S.-K. Park, H., Kim, W., S. Park, J., H. Jeong, emn,G . ole,C . olc,S . ’ogln .W,Gpa S., Gupta, W., C. O’Loughlin, J., S. Pollock, F., C. Woeller, M., G. Lehmann, Fromigue J.-P., Marolleau, S., Lecourt, anr . nemn,G,Wbtc,M,Gilr,D,Alad . erl .and R. Negrel, G., Ailhaud, D., Gaillard, M., Wabitsch, G., Entenmann, H., Hauner, Ja C. M. Dalakas, and M. Leon-Monzon, I., Illa, M. W. B. E. Spiegelman, Kraegen, and and P. J. Tontonoz, G. E., Hu, Cooney, J., Ye, M., S. Furler, D., B. Hegarty, e,J . uPtre,Z,Co . iua . akwk,R,Cmis . Usas, J., Cummins, R., Jankowski, S., Kimura, B., Cao, Z., Qu-Petersen, Y., J. Lee, ¨a gr .adNua,T. Neuman, and K. ¨ger, oncietsu eeomn n rvsssei fibrosis. systemic drives and development tissue connective inflammation. musculoskeletal and disease Pharmacol. metabolic Opin. in adipokines muscle: 384. 401. V. myogenesis. facilitate M. that F. progenitors Rossi, and 20) dpgnssadaig osaigmk a oMAD? go fat make aging does 757-767. aging: and Adipogenesis (2002). differentiation. 21) aelt el,cnetv isefbolssadteritrcin r crucial are interactions their and regeneration. fibroblasts muscle tissue for connective cells, Satellite (2011). G. Kardon, and 493-495. M. Angus-Hill, myogenesis. S., regulate Tcf4 M. and fibroblasts Hansen, tissue A., Connective D. Hutcheson, individuals. P. biopsies. muscle skeletal F. human Kadi, and L. Jfbols el niisteaioeei n ioeei n33L cells. 3T3-L1 in Commun. lipogenesis and Res. adipogenesis Biophys. the inhibits cells fibroblast BJ edn .E n hps .P. R. fibroblasts. Phipps, human and E. S. Feldon, activation. fibroblast in partners PDGF, and Res. CCN2, endothelin, angiotensin, quantification automated lipids. and of immunofluorescence with combination permits staining ua dpct rcro el utrdi hmclydfndmedium. defined chemically a in cultured Invest. cells and precursor adipocyte acids human fatty F. of E. Pfeiffer, effects triggering adipoblasts: to thiazolidinediones. myoblasts of differentiation kltlmsl erlcl deinmlcl NCM:cagsi rti and protein in changes lines. cell (N-CAM): muscle molecule of myogenesis adhesion in during species cell regeneration mRNA neural muscle muscle skeletal Skeletal to cells mice. muscle-derived host human dystrophic of Contribution fibrosis. regeneration. muscle muscle skeletal skeletal injury-induced acute in Development cells satellite Pax7-positive dpgncdfeetaincmae ihmsnhmlse cells. stem mesenchymal 20 with compared differentiation adipogenic epithelial endocrine and microenvironment. mesodermal-derived thymic human of the of ontogeny components and characterization eie el aal fehnigmsl eeeainadbn healing. al. bone and et regeneration A. muscle 150 enhancing Wernig, of capable P., cells Robbins, derived C., Gates, A., Boume I., Robert, M.-N., Lacassagne, B., Ternaux, ua ucefbr n aelt el xrs erlcl deinmolecule adhesion cell neural express cells. killer cells natural to satellite antibodies monoclonal and by fibers recognized muscle human alpha. C/EBP and gamma USA PPAR Sci. factors Acad. transcription adipogenic the by resistance. insulin in lipid intramuscular of role kltlmsl nst n nvitro. human in from and populations situ cell in mesenchymal-like muscle skeletal distinct of Characterization (2010). 1327-1336. , 21) nrmsua dps ise acpna n oiiyfnto nolder in function mobility and sarcopenia, tissue, adipose Intramuscular (2012). 1085-1100. , 106 84 1675-1680. , 16) aelt elo kltlmsl fibers. muscle skeletal of cell Satellite (1961). itce.Cl Biol. Cell Histochem. 1663-1670. , 21) oeta ther Potential (2010). .AigRes. Aging J. 138 hso.Rev. Physiol. 18) rmtn feto lccriod ntedfeetainof differentiation the on glucocorticoids of effect Promoting (1989). 92 3639-3646. , 20) seseto aelt elnme n ciiysau in status activity and number cell satellite of Assessment (2009). 4 9856-9860. , m .Physiol. J. Am. rsalnisLuo.Esn.FtyAcids Fatty Essent. Leukot. Prostaglandins 281-289. , LSONE PLoS 392 21) uceijr ciae eietfibro/adipogenic resident activates injury Muscle (2010). .Bo.Chem. Biol. J. Development 2012 520-525. , 20) irbat ncancer. in Fibroblasts (2006). 78 21) ua emlfbolssehbtdelayed exhibit fibroblasts dermal Human (2011). 629637. , 20) nrae DFapaatvto disrupts activation PDGFRalpha Increased (2009). 783-809. , 116 uceNerve Muscle 21) oe niaioei ciiypoue by produced activity anti-adipogenic Novel (2010). pui agt o ada irss TGFbeta, fibrosis: cardiac for targets apeutic 6 299 e17454. , 63-68. , x.Cl Res. Cell Exp. 20) h eiaeblnebtenftand fat between balance delicate The (2004). ,O,Vuhz . nraaaian,R., Andriamanalijaona, K., Vauchez, O., ´, 138 a.Cl Biol. Cell Nat. C672-C681. , 283 3625-3637. , 19) rndfeetaino myoblasts of Transdifferentiation (1995). 19371-19378. , 21) naslt eurmn for requirement absolute An (2011). 19) eeeaigaddenervated and Regenerating (1992). 20) ysai ietyregulates directly Myostatin (2008). 40 caPyil Scand. Physiol. Acta 20) lnlioaino muscle- of isolation Clonal (2000). 455-465. , 20) piiaino i e O red oil of Optimisation (2001). 19) nesadn adipocyte Understanding (1998). 316 dee . Che K., ´diene, .Ep Med. Exp. J. 12 .Bohs ice.Cytol. Biochem. Biophys. J. 2513-2526. , .Cl Biol. Cell J. 153-163. , a.Rv Cancer Rev. Nat. n.Neurol. Ann. e.Cell Dev. Development 21) outr with Coculture (2010). 57 18) Phenotypic (1984). x.Gerontol. Exp. 71-75. , 159 105 tmClsDev. Cells Stem ra,F tal. et F. ´reau, 178 19) Trans- (1997). 16 1149-1168. , 1377-1386. , .Cl Biol. Cell J. 20) The (2003). rc Natl. Proc. 303-313. , 373-383. , 31 138 Biochem. 46-52. , .Clin. J. 6 (2011). (1987). (2011). 392- , 371- , Curr. Circ. 37 9 , , Journal of Cell Science aael,P,Bnlo . atni . rget,P,Brai,N,Cpni C., Capanni, N., Bergamin, P., Braghetta, G., Lattanzi, P., Bonaldo, P., Sabatelli, usl,T .adH,R. Ho, and R. T. Russell, ui,C . ish . ug .adRsn .M. O. Rosen, and C. Fung, A., Hirsch, S., C. Rubin, M. K. Yamada, and M. Rossner, Ramı S., Merfeld-Clauss, H., B. Johnstone, C., W. Roell, O., D. Traktuev, G., Rajashekhar, oe,E . s,C-. ag . aa,S,Femn .W,Gnae,F .and J. F. Gonzalez, W., M. Freeman, S., Sakai, X., Wang, C.-H., Hsu, D., E. Rosen, M. B. Spiegelman, and D. E. Rosen, R. S. Smith, and E. Ravussin, Randolph- R., Pruchnic, J., T., Cummins, M., Goodpaster, Ikezawa, R., M., Jankowski, B., Deasy, E. Z., Qu-Petersen, Haley, A., Legesse-Miller, A., J.-Y., Kurzenne, E. S., Sacconi, Pollina, M., Plaisant, A., Loubat, N., Clement, F., D. Pisani, iln,D,Fn . un,D,Ku,B n oe,R H. R. Gomer, and B. Kaul, D., Huang, T., Fan, D., Pilling, hfr . lkisiLe .adYbok-evn,Z. Yablonka-Reuveni, and M. Wleklinski-Lee, G., Shefer, A. M. Lazar, and D. Shao, fsee,U,Krey . opr . od . eadr . uor A., Dufour, J., Nelander, J., and Wood, C. O., McAllister, Torper, K., A., E. Kirkeby, Johnson, U., M., Pfisterer, J. Thomas-Ahner, M., C. Penton, er,D . sa,G,Hio . u .G,Cute,C . fei,J M., J. Ofrecio, H., C. Courtney, G., J. Yu, A., Hsiao, G., Hsiao, D., A. D. Starzinski-Powitz, Sears, and M. Cramer, K., Zimmermann, W., Schubert, D. Yaffe, and U. Nudel, A., Panski, L., Tencer, A., O., Fuchs, Paldi, R., Sarig, L., Wittenberghe, Van A., Kissenpfennig, R., Yao, R., Sambasivan, crue-idrig .B,Hseik .K . crue,P n oi .E. M. Kooi, and P. Schrauwen, C., K. M. Hesselink, B., V. Schrauwen-Hinderling, atoi . oubr,M,Onbn,A,Pp,G tal. et G. Pepe, A., Ognibene, M., Columbaro, E., Mattioli, rgeigo ifrnito ypotgadnFapaad1-methyl-3-isobutyl and F2alpha prostaglandin by differentiation xanthine. of triggering eetr n omnlrsosvns nvto nui eetr n insulin and receptors cells. 3T3-L1 Insulin of vitro. forms adipocyte in and 253 preadipocyte responsiveness the in hormonal sensitivity and receptors signaling. Wnt paracrine and canonical contact of cell endothelial role M. by communication: Clauss, reduced and is L. differentiation K. cell March, stromal Adipose D., E. Rosen, B., Natta, Van manipulation. pathway. unified a M. B. Spiegelman, type and resistance, insulin storage, fat mellitus. ectopic in diabetes result 2 proliferation cell fat intracytoplasmic of staining failure by O. triglycerides red and Oil with conversion lipids adipose of Quantitation regeneration. muscle for potential mice: in cells al. et stem Biol. A. muscle Wernig, of C., Gates, population B., Cao, J., Mytinger, Cycle A. Cell H. Coller, and J. Habecker, muscle. skeletal A. human C. within Dechesne, progenitors and adipogenic and C. Dani, C., Desnuelle, n lcs homeostasis. glucose and macrophages, monocytes, fibroblasts. from fibrocytes and monocyte-derived distinguish that markers aelt el a pnaeul ne natraiemsnhmlpathway. mesenchymal alternative an enter Sci. spontaneously can cells satellite ovrino ua irbat odpmnri neurons. dopaminergic 108 to fibroblasts human of conversion Bjo fate. fibro-adipogenic a adopt muscle F. Montanaro, dpct differentiation. adipocyte protein CCAAT/enhancer-binding sensitization. S. insulin USA Sci. thiazolidinedione-mediated Subramaniam, and and resistance J. Chapman, regeneration. muscle skeletal A. adult of Galy, Development for and indispensable are S. matrix cells Tajbakhsh, satellite B., extracellular Pax7-expressing Malissen, H., the Guenou, B., in Gayraud-Morel, fibronectin of organization fibroblasts. the cultured affects deficiency ypoyeatgnLu1 samlclrmre frgnrto nhmnskeletal human in regeneration of marker molecular muscle. a as Leu-19 antigen muscle. Lymphocyte skeletal human in content Spring) lipid Intramyocellular cells. (2006). neuron-like into vivo in transdifferentiate e8814. can cells myogenic ´rez-Zacarı rln,A,Lnvl,O,Jkbsn .adPra,M. Parmar, and J. Jakobsson, O., Lindvall, A., ¨rklund, 7570-7578. , 10343-10348. , 117 157 5393-5404. , 14 rc al cd c.USA Sci. Acad. Natl. Proc. 851-864. , 106 rc al cd c.USA Sci. Acad. Natl. Proc. 357-367. , 7 2056-2070. , a,J . Castro-Mun L., J. ´as, 18745-18750. , 138 .Cl Biol. Cell J. LSONE PLoS 21) ucesd ouainclsfo ytohco injured or dystrophic from cells population side Muscle (2013). 3647-3656. , ee Dev. Genes 20) /Bapaidcsaioeei hog PPARgamma: through adipogenesis induces C/EBPalpha (2002). n.N .Aa.Sci. Acad. Y. N. Ann. arxBiol. Matrix Histochemistry .Bo.Chem. Biol. J. 17) ovrino T irbat noaioecells: adipose into fibroblasts 3T3 of Conversion (1976). Nature 166 4 20) nrae a nae mardftoiain and oxidation, fat impaired intake, fat Increased (2002). e7475. , 19) eoioepoieao ciae receptor activated proliferator Peroxisome (1997). 11-15. , 16 20) htsi itr?Tetmtto fimage of temptation The picture? a in What’s (2004). 20) euaigteagoei aac ntissues. in balance angiogenic the Regulating (2008). 20 444 22-26. , a 20) dpctsa euaoso nrybalance energy of regulators as Adipocytes (2006). n elccesau euaetecmimn to commitment the regulate status cycle cell and , 475-486. , 86 847-853. , old,F n uiHruh W. Kuri-Harcuch, and F. ˜ozledo, 73 97 LSONE PLoS 307-311. , 4516-4520. , 493-497. , 272 967 20) ehnsso ua insulin human of Mechanisms (2009). 21473-21478. , 363-378. , 21) irrhzto fmyogenic of Hierarchization (2010). tmCells Stem 8 e54553. , 17) eeomn fhormone of Development (1978). 20) dniiaino novel a of Identification (2002). rc al cd c.USA Sci. Acad. Natl. Proc. tmCells Stem 20) FT collection: IFATS (2008). 26 20) dniiainof Identification (2009). 20) kltlmuscle Skeletal (2004). 2674-2681. , 20) olgnVI Collagen (2001). rc al Acad. Natl. Proc. bst (Silver Obesity 28 21) Cloned (2010). 21) Direct (2011). .Bo.Chem. Biol. J. LSONE PLoS 2182-2194. , (1992). (1989). (2011). .Cell J. .Cell J. c 5 , , amt .S,Prrde .A n alnaRuei Z. Yablonka-Reuveni, and A. T. Partridge, S., P. Zammit, R. J. Beauchamp, and S. P. Zammit, i,J . eha . lu . emfed . una . ars .and T. Harris, P., A. Kuznia, M. Rudnicki, S., and F. Heymsfield, Price, H., J., Yin, Albu, S., Heshka, E., J. Yim, at .J,Hy .J,Moo .M n oea,R A. R. Coleman, and M. D. Muoio, J., A. Hoy, J., M. Watt, C. J. Waters, a,D . h,Y-. aaui .P,Qat,N,Los .M n Longaker, and M. K. Lyons, N., Quarto, P., R. Nacamuli, Y.-Y., Shi, C., D. Wan, aasm,A. Hashimoto, and Wagatsuma, S. Yasumoto, S., Shimizu, M., Inagawa-Ogashiwa, R., M. Wada, etr . ia,G,Fazn . an,M,D op,P,Rzuo .and R. Rizzuto, P., Coppi, De M., Sanna, C., Franzin, G., Milan, R., Vettor, etn,A . alrJns .M,Lno .A,Badn .D,Ln,T F., T. Lane, D., E. Bearden, A., K. Longo, M., J. Taylor-Jones, M., A. Vertino, a e e,P . car,G,Jp .H,Snes .C,Sahues .M and M. A. Stadhouders, C., R. Sengers, H., P. Jap, G., Schaart, F., P. Ven, der van ckna . iraz,N,Prshlv,T,Too,T,DPne . o,A., Koo, M., DePonte, T., Thomou, T., Pirtskhalava, N., Giorgadze, T., Tchkonia, eui . t,T,Mrkw,D,Siiu . oea . eaa M., Segawa, T., Yoneda, N., Shimizu, D., Morikawa, T., Ito, A., Uezumi, ag .Q n ae .D. M. Lane, and Q. D. Q. M. Tang, Lane, and Y. R. Hsu, K., A. Student, eui . uaa .i,Ymmt,N,Tkd,S .adTuhd,K. Tsuchida, and i. S. Takeda, N., Yamamoto, S.-i., Fukada, A., Uezumi, twr,J . ai .L,Cmig .E,Mla,G . etot,B M., B. Wentworth, M., G. Molnar, E., A. Cumming, L., T. Masi, D., J. Stewart, ye,K L. K. Tyler, o,Y,Zag .Z,Sbtli . h,M-.adBo and M.-L. Chu, P., Sabatelli, R.-Z., Zhang, Y., Zou, tre,J . aaoo . aaoo .adGlhmr .J. D. Goldhamer, and S. Yamamoto, M., Yamamoto, D., J. R. Starkey, S. Farmer, and M. B. Spiegelman, ute .J,Sncr,D .adMelr .J. M. Mueller, and R. D. Sinacore, J., L. Tuttle, hn,B,Co . rsn . u,B,L,G,Lgr . a,S,Plet .B., J. Pollett, S., Yap, A., Logar, G., Li, B., Sun, M., Crisan, B., Cao, B., Zheng, ucestliecl:tese elta aei rmtecold. the from in came that cell stem the cell: satellite muscle cell? stem of son or cell niche. risk. cardiovascular with associations independent D. Gallagher, pcfcftyaisi ellrpoess mlctosfritrrtn n reporting and interpreting for implications processes: cellular experiments. in acids fatty specific eursrtni cdadbn opoeei rti eetrtp Bsignaling. Biol. IB Cell type J. receptor protein USA morphogenetic Sci. Acad. bone Natl. and Proc. acid retinoic requires T. M. regeneration. Development N. ahpyilgclimplications. pathophysiological G. Federspil, eiinypooe oxrsino ygncadaioei rgasin programs adipogenic A. and C. myogenic Peterson, and of A. coexpression O. myoblasts. MacDougald, promotes Jr, deficiency E., R. McGehee, aakr,F C. F. Ramaekers, auaina niae yttnaddsi striation. desmin and titin by indicated as maturation eo-pcfccaatrsisaertie nsrisdrvdfo igehuman single from derived strains in al. retained et T. are preadipocytes. Zglinicki, von characteristics C., Martin-Ruiz, depot-specific D., Chinnappan, A., R. Forse, irssadaioeei rgnt rmacmo eecya rgntrin progenitor mesenchymal al. common et a Y. from Miyagoe-Suzuki, originate M., muscle. skeletal M. adipogenesis Matev, and R., Fibrosis Ogawa, M., Yamaguchi, e.Biochem. Rev. ytei ndfeetaig33L preadipocytes. 3T3-L1 differentiating in synthesis of modulation C. differential TGF- P. vitro: by in Yaeger, markers cells myoblast and muscle-derived M. skeletal human J. proliferating McPherson, K., Sampath, eecya rgntr itntfo aelt el otiuet coi a cell fat ectopic to contribute muscle. cells skeletal satellite in from formation distinct progenitors Mesenchymal Nerve Muscle ucestlieclsaecmitdt ygnssadd o pnaeul adopt spontaneously not do and myogenesis to fates. committed nonmyogenic are cells satellite muscle smsl pcfcadascae ihpo ucinlperformance. functional poor with associated and 2012 specific muscle is uce mlctosfrcneia uclrdsrpytpsUlihadBethlem. and Ullrich skeletal types Neurol. in dystrophy Exp. synthesis muscular Neuropathol. VI J. collagen congenital of for source implications main the muscle: are fibroblasts interstitial Muscle muscle. al. skeletal et human in T. cells endothelial Cassino, L., Drowley, Cytochem. xrsinpirt opooia ifrnito f33adipocytes. 3T3 of differentiation morphological to prior expression rndfeetaino ucefbolss5625 fibroblasts muscle of Transdifferentiation 20) eeaino ifrn ae rmmlioetmsl tmcells. stem muscle multipotent from fates different of Generation (2002). 172957. , 20) segncdfeetaino os dps-eie dl toa cells stromal adult adipose-derived mouse of differentiation Osteogenic (2006). hso.Rev. Physiol. 20) rgn n al ecitoso ‘uhnemsua dystrophy’’. muscular ‘‘Duchenne of descriptions early and Origins (2003). 54 o.Bo.Cell Biol. Mol. 20) cuayadpeiini uniaiefursec microscopy. fluorescence quantitative in precision and Accuracy (2009). 185 m .Physiol. J. Am. 1177-1191. , o.Cl.Biochem. Cell. Mol. 129 28 Diabetes 81 20) nemsua dps iservl icrlaioetsu in tissue adipose visceral rivals tissue adipose Intermuscular (2007). 20) dpgncptnilcnb ciae uigmuscle during activated be can potential Adipogenic (2007). 1135-1148. , .Cl Sci. Cell J. 20) h rgno nemsua dps iseadits and tissue adipose intermuscular of origin The (2009). 402-422. , 2987-2995. , 715-736. , 19) ifrnito fhmnseea uceclsi culture: in cells muscle skeletal human of Differentiation (1992). .Hsohm Cytochem. Histochem. J. 93 23-67. , 55 Differentiation 2571-2578. , 16 b 21) dpgnss rmse elt adipocyte. to cell stem from Adipogenesis: (2012). 103 2. 124 302 2039-2048. , 67 a.Cl Biol. Cell Nat. .Cl.Physiol. Cell. J. 12335-12340. , 3654-3664. , 144-154. , m .Physiol. J. Am. E1-E3. , 304 20) rsetv dniiaino myogenic of identification Prospective (2007). 20) h kltlmsl aelt el stem cell: satellite muscle skeletal The (2001). 21) aelt el n h ucese cell stem muscle the and cells Satellite (2013). 25-33. , 18) erae ntblnadatngene actin and tubulin in Decreases (1982). 68 193-204. , 18) nuto fftyai synthetase acid fatty of Induction (1980). a.Biotechnol. Nat. 12 59 143-152. , 196 33-46. , 21) nemsua dps tissue adipose Intermuscular (2012). 297 n.J Obes. J. Int. .Bo.Chem. Biol. J. 70-78. , E987-E998. , elTsu Res. Tissue Cell 20) hrceiainof Characterization (2003). nean .G. C. ¨nnemann, 21) itntrlsof roles Distinct (2012). 25 (Lond) 1025-1034. , 20) h skeletal The (2006). 255 20) Wnt10b (2005). 21) Skeletal (2011). Cell 31 4745-4750. , .Histochem. J. 270 .AigRes. Aging J. 1400-1405. , 20) Fat (2006). 29 189-198. , 53-60. , (2011). (2010). (2008). Annu.