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Stem Cell Quick Guide Stem Cells by Miltenyi Biotec Quick guide inside! miltenyibiotec.com/stemcells Embryonic and induced pluripotent stem cells Pioneering solutions for ES and iPS cells research Differentiation Reprogramming Purification of ES and iPS cells prior to differentiation Fibroblasts isolation Human Mouse • Whole Skin Dissociation Kit • TRA-1-60 MicroBeads • Pluripotent Stem Cell • gentleMACS™ Dissociators • Feeder Removal Isolation Kit MicroBeads • Feeder Removal • Anti-Fibroblast MicroBeads Generation of reprogrammed cells MicroBeads • StemMACS mRNAs for reprogramming and differentiation Induction of differentiation • StemMACS mRNA Transfection Reagent • StemMACS Small Molecules • MACS Cytokines • StemMACS™ Small Molecules, such as CHIR99021, Purmorphamine, and more Isolation of reprogrammed cells • StemMACS mRNA, including Brn2, Mash1 Human Mouse Note: Different reagents are used in accordance • Anti-TRA-1-60 • Anti-SSEA-1 MicroBeads with the differentiation pathway. MicroBeads • Flow sorting using • Anti-SSEA-4 MicroBeads the MACSQuant Tyto Transfection and transduction of ES and iPS cells • StemMACS Y27632 • StemMACS Thiazovivin • MACSelect™ Products • Flow sorting using • StemMACS mRNA Transfection Kit the MACSQuant Tyto • StemMACS Nuclear eGFP mRNA • MACSfectin™ Monitoring differentiation by flow cytometry Maintenance or immunohistochemistry Culture of ES and iPS cells • MACS Antibody Portfolio Example application: Use CD34 antibodies Human Mouse to analyze cells during differentiation into • StemMACS iPS-Brew XF • MACS Cytokines, such hematopoietic or endothelial lineages. • StemMACS Passaging as LIF • MACSQuant® Flow Cytometers Solution XF • StemMACS™ Small • StemMACS Small Molecules, such as Purification of cells during or after differentiation Molecules, such as CHIR99021, PD0325901 Y27632, Thiazovivin, Removal of remaining ES or iPS cells CHIR99021, PD0325901 • UEA-1 MicroBeads • MACS Cytokines, such as FGF-2 Ectoderm • Anti-TRA-1-60 MicroBeads • Neural Crest Stem Cell MicroBeads • Anti-A2B5 MicroBeads Kit • Anti-PSA-NCAM MicroBead Kit Phenotyping of ES and iPS cells • CD133 MicroBead Kit Endoderm Human Mouse • CD184 (CXCR4) MicroBead Kit MACSQuant Flow MACSQuant Flow Cytometers and Cytometers and Mesoderm fluorochrome-conjugated fluorochrome- • CD34 MicroBead Kit antibodies for detection of conjugated antibodies for • CD144 MicroBeads • Pluripotency: TRA-1-60, detection of: • CD309 (Flk-1) MicoBead Kit TRA-1-81, SSEA-4, • Pluripotency marker, CD326, CD324 such as SSEA-1, CD326 • Differentiated cells: • Feeder cells: SSEA-1 Anti-Feeder • Feeder cells: Anti-Feeder, Anti-Fibroblast Sample preparation Cell separation Flow cytometry Cell culture Molecular applications Clinical applications Hematopoietic stem cells Making headway in your HSC research Sample preparation • Dead Cell Removal Kit • Smart Strainers • Pre-Separation Filters • Red Blood Cell Lysis Solution Human samples Mouse samples Isolation of HSCs Isolation of HSCs • CD34 MicroBead Kit UltraPure • Lineage Cell Depletion Kit • CD133 MicroBead Kit – Hematopoietic Tissue • Anti-Sca-1 MicroBead Kit (FITC) • CD117 MicroBead Kit • CD117 MicroBeads • Lineage Cell Depletion Kit • CD105 MultiSort Kit (PE) • Flow sorting using the MACSQuant Tyto and your favorite • Flow sorting using the MACSQuant Tyto and your favorite marker panel marker panel Culture of HSCs Culture of HSCs Expansion Expansion • StemMACS HSC Expansion Media XF • MACS Cytokines, including SCF, Flt3-Ligand, Il-3, Il-6 • StemMACS HSC Expansion Cocktail Differentiation • MACS Cytokines, including TPO, SCF, Flt3-Ligand, Il-3, Il-6 • MACS Cytokines, such as Il-7, GM-CSF Differentiation • StemMACS HSC-CFU Media • MACS Cytokines, such as Il-7, GM-CSF Phenotyping and flow cytometry Phenotyping and flow cytometry MACSQuant Flow Cytometers in combination with MACSQuant Flow Cytometers in combination with fluorochrome-conjugated MACS Antibodies including: fluorochrome-conjugated MACS Antibodies, such as • CD34 • Lineage markers • CD133 • Sca-1 • CD45 CHECK OUT OUR • CD117 (c-kit) • CD38 EXCLUSIVE VIO®DYES! • CD135 (Flt3) • CD90 Molecular anaylsis of HSCs Translate research to clinical applications • MACS GMP Cytokines • µMACS™ mRNA Isolation Kit • SuperAmp™ Technology – cDNA • CliniMACS® and CliniMACS Prodigy® • µMACS Protein Isolation Kits from just a few cells • CliniMACS Reagents, including CD133 (epitope tags, protein A/G) • Magnetic mitochondria isolation • MultiMACS™ M96 Separator • Enhanced viral transduction with MACSductin StemMACS HSC Expansion Media XF • Xeno-free expansion of hematopoietic stem and progenitor cells • Consistently high expansion rates • High CD34+ cell content after expansion Mesenchymal stem cells Driving your MSC research Sample preparation • Dead Cell Removal Kit • Smart Strainers • Pre-Separation Filters • gentleMACS Dissociators Human samples Mouse samples Phenotyping or flow sorting of MSCs prior to culture Phenotyping or flow sorting of MSCs prior to culture Fluorochrome-conjugated antibodies against, for example: Fluorochrome-conjugated antibodies against, for example: • CD271 • CD146 • Lineage markers • CD45 • MSCA-1 • CD105 • Sca-1 • CD105 Isolation of MSCs prior to culture Isolation of MSCs prior to culture MicroBeads for positive isolation MicroBeads for removal of non-MSCs • CD271 MicroBead Kit • CD146 MicroBead Kit • Lineage Cell Depletion Kit • Anti -Ter119 MicroBeads • Anti-MSCA-1 (W8B2) • CD105 MicroBeads • CD45 MicroBeads • CD11b MicroBeads MicroBead Kit • Anti-Fibroblast MicroBeads Culture of MSCs Culture of MSCs Expansion Differentiation • MACS Cytokines, including TGF-β, EGF • StemMACS MSC • StemMACS AdipoDiff Expansion Media Kit XF • StemMACS ChondroDiff • StemMACS MSC • StemMACS OsteoDiff Phenotyping of expanded MSCs Expansion Media • CytoMix – MSC • Fluorochrome-conjugated antibodies against, for example, • MACS Cytokines, such as lineage markers, CD105, CD117, Anti-Sca-1, CD45, CD116, FGF-2, PDGF-BB, TGF-β and more Phenotyping of expanded MSCs • MSC Phenotyping Kit • Fluorochrome-conjugated antibodies against, for example, CD73, CD90, CD105, CD45, CD34, CD14, CD20, and more Molecular anaylsis of MSCs Translate research to clinical applications • MACS GMP Cytokines • µMACS mRNA Isolation Kit • SuperAmp™ Technology – cDNA • CliniMACS and CliniMACS Prodigy • µMACS Protein Isolation Kits from just a few cells • CliniMACS CD271 Reagent (epitope tags, protein A/G) • Magnetic mitochondria isolation • MultiMACS™ M96 Separator • Enhanced viral transduction with MACSductin™ StemMACS MSC Expansion Media Kit XF • Xeno-free expansion of human mesenchymal stem cells • No attachment substrate required • Excellent expansion rates Embryonic and induced pluripotent stem cells Highlighted products StemMACS iPS-Brew XF • Xeno-free culture system for human ES and iPS cells • Easy transition to feeder-free conditions • Formulation compliant with future clinical translation Learn more StemMACS™ Small Molecules • Highly pure for consistent biological activity • Pre-tested on stem cell cultures • Detailed instructions for preparation of stock solutions Request your free poster StemMACS™ mRNA Technology • Modified mRNAs for modulation of cell fate by mRNA transfection • Optimized sequences, validated expression • Validated protocol for mRNA reprogramming Download the mRNA reprogramming manual Human FGF-2 IS • Improved sequence for increased stability in cell culture • Standardized, high biological activity • Produced in E. coli, free of animal components See the data Fore more information visit www.miltenyibiotec.com/stemcells Germany/Austria/ Benelux France Nordics and Baltics South Korea Switzerland Miltenyi Biotec B.V. Miltenyi Biotec SAS Miltenyi Biotec Norden AB Miltenyi Biotec Korea Miltenyi Biotec GmbH Schipholweg 68 H 10 rue Mercoeur Scheelevägen 17 Petra building Friedrich-Ebert-Straße 68 2316 XE Leiden 75011 Paris, France 223 70 Lund 659 Yeoksam-dong 51429 Bergisch Gladbach The Netherlands Phone +33 1 56 98 16 16 Sweden Gangnam-gu Germany [email protected] Fax +33 1 56 98 16 17 [email protected] Seoul, South Korea Phone +49 2204 8306-0 Customer service [email protected] Customer service Sweden Phone +82 2 555 1988 Fax +49 2204 85197 The Netherlands Phone 0200 -111 800 Fax +82 2 555 8890 [email protected] Phone 0800 4020120 Italy Fax 046-280 72 99 [email protected] Fax 0800 4020100 Miltenyi Biotec S.r.l. Customer service Denmark USA/Canada Customer service Belgium Via Persicetana, 2/D Phone 80 20 30 10 Spain Miltenyi Biotec Inc. 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