ORGANOIDS Robust Media to Culture Intestinal, Hepatic, Pancreatic, Kidney, Airway, and Cerebral Organoids
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2 STEMCELL Technologies Inc. TABLE OF CONTENTS
4 Introduction
5 Intestinal Organoids 6 IntestiCult™ Organoid Growth Medium (Mouse) 7 IntestiCult™ Organoid Growth Medium (Human) 8 IntestiCult™ Organoid Differentiation Medium (Human) 11 STEMdiff™ Intestinal Organoid Kit
13 Hepatic Organoids 13 HepatiCult™ Organoid Growth Medium (Mouse)
16 Pancreatic Organoids 16 PancreaCult™ Organoid Growth Medium (Mouse)
19 Kidney Organoids 19 STEMdiff™ Kidney Organoid Kit
22 Airway Organoids 22 PneumaCult™ Airway Organoid Kit
24 Cerebral Organoids 24 STEMdiff™ Cerebral Organoid Kit
26 Product Information & References
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Introduction
One of the most impactful advances in stem cell research of Removal of confounding variables inherent in animal models while the past decade has been the development of organoid culture providing increased complexity compared with homogeneous techniques. Organoids are three-dimensional cell cultures that cell cultures gives organoid cultures specific experimental recapitulate some of the key cell types and structural features advantages. By combining a high level of physiological relevance of the represented organ. These “mini-organs” enable scientists with amenability to manipulation in vitro, organoids have the to investigate complex research questions in vitro by providing potential to complement or replace in vitro experiments using highly relevant tissue model systems. Since landmark publications primary cells or immortalized cell lines. There is also a movement describing intestinal1 and cerebral organoids,2 organoid culture to use organoids to replace animal experimentation in many techniques have been developed for multiple species and tissues to circumstances. Additionally, organoids demonstrate high genetic derive organoids from either adult stem cells (ASCs) or pluripotent stability in culture, retaining the genotype and phenotype of the stem cells (PSCs). source tissue. Organoids therefore provide faithful models of disease and have been used to model disease mechanisms and While each culture system is distinct, organoids generally contain progression, as well as to predict patient-specific response to drug multiple different organ-specific cell types capable of recapitulating treatment. some functions of that organ. During organoid culture, stem cells (PSCs or ASCs) are provided with appropriate signaling factors Continued development of organoid methodologies is proving that mimic the in vivo environment. Exposure to these conditions transformative to the stem cell research landscape. In many labs, directs stem cells to divide and differentiate into the cell types of organoids have complemented model systems already in place, the represented organ. In general, organoids display architecture while in others they have enabled investigations previously not similar to what is observed in vivo, such as polarization of the possible. Development and standardization of organoid culture apical and basolateral surfaces (epithelial organoids), appropriate techniques will enable even greater accessibility of organoids. cellular stratification (cerebral organoids), or re-creation of complex domains such as the intestinal crypt-villus structure observed in mouse intestinal organoids. In epithelial organoid systems, the presence of actively dividing adult stem cells enable the cultures to be expanded and maintained in long-term culture through serial passaging. Organoid cultures have seen quick and widespread adoption for a variety of applications, from basic research to translational and industrial uses. The roots of organoid technology lie in developmental biology, and organoids are now feeding back valuable information about the development of the tissues that they model, especially in the case of PSC-derived organoids. As organoids recapitulate many of the signaling and cell-cell interactions observed in vivo, they are also invaluable models for studying cell biology, including investigation into tissue regeneration mechanisms, maintenance of the stem cell niche, and interactions with bacteria, viruses, and cells from other tissues.
4 STEMCELL Technologies Inc. Intestinal Organoids
Intestinal organoids incorporate key intestinal cell types including long‑term maintenance of ASC-derived intestinal organoids enterocytes, goblet cells, enteroendocrine cells, transit amplifying from mouse and human cells, respectively. STEMdiff™ Intestinal cells, and intestinal stem cells, recapitulating the cellular Organoid Kit can be used to direct human PSCs through a three- complement of the intestinal epithelium. The presence of an actively stage differentiation process to intestinal organoids that can dividing stem cell population enables expansion and maintenance of be maintained and matured in STEMdiff™ Intestinal Organoid intestinal organoids in long-term culture. Growth Medium. Intestinal organoids grown using these media are convenient, flexible, and relevant intestinal tissue models that can IntestiCult™ Organoid Growth Medium (Mouse) and IntestiCult™ increase experimental impact and reduce laboratory animal use. Organoid Growth Medium (Human) support establishment and
Table 1. Comparison of Intestinal Organoid Culture Systems
INTESTINAL ORGANOID TYPE MOUSE (ASC-DERIVED) HUMAN (ASC-DERIVED) HUMAN (PSC-DERIVED)
Representative Image
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Organoid Cellular Composition Organoids simultaneously After 1 - 3 passages organoids Organoids simultaneously incorporate an active intestinal are composed primarily of cells incorporate an active intestinal stem cell compartment and in a progenitor/stem-like state; stem cell compartment, differentiated cell types in an organoids can be differentiated differentiated intestinal epithelium that recapitulates the to mature intestinal cells in an epithelial cell types and in vivo intestine. appropriate medium. associated mesenchymal cells. Organoids are phenotypically fetal.
Starting Material • Mouse intestinal or • Human intestinal or • Human induced pluripotent colonic crypts colonic crypts stem cells (iPSC) or embryonic • Mouse Intestinal Organoids • Established ASC-derived stem cell (ESC) lines (Catalog #70931) intestinal organoids • Established PSC-derived intestinal organoids
Organoid Maintenance Organoids can be maintained Organoids can be maintained Organoids can be maintained through long-term passaging through long-term passaging through long-term passaging or cryopreserved. or cryopreserved. or cryopreserved.
Genetic Considerations • Established common healthy • Donor-specific genetic • Donor-specific genetic and disease model strains background background • Tools for in vivo genetic • Targeted in vitro gene editing • Targeted in vitro gene editing manipulation possible possible
Compatible Culture Media IntestiCult™ Organoid IntestiCult™ Organoid STEMdiff™ Intestinal Growth Medium (Mouse) Growth Medium (Human) Organoid Kit (Catalog #05140) (Catalog #06005) (Catalog #06010) STEMdiff™ Intestinal IntestiCult™ Organoid Organoid Growth Medium Differentiation Medium (Catalog #05145) (Human) (Catalog #100- 0214); for differentiation in 3D organoids and as monolayers in submerged or air-liquid interface cultures.
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IntestiCult™ Organoid Why Use IntestiCult™ Organoid Growth Medium (Mouse) Growth Medium (Mouse)? Cell Culture Medium for RELEVANT. Enables generation of intestinal organoid cultures that recapitulate the identity and organization of Establishment and Maintenance the adult intestinal epithelium.
of Mouse Intestinal Organoids ROBUST. Supports efficient establishment of organoids from mouse intestinal crypts in less than one week. IntestiCult™ Organoid Growth Medium (Mouse) is a complete, serum‑free organoid growth medium that enables researchers to SIMPLE. Convenient format and easy-to-follow protocol. easily and reproducibly generate experiment-ready organoids in SERUM-FREE. Optimized formulation for low 5 - 7 days. experimental variability.
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Figure 2. IntestiCult™ Organoid Growth Medium (Mouse) Supports Efficient Organoid Establishment and Expansion
(A) Established organoids can be passaged efficiently over an indefinite number of passages. (B) Starting from a single well containing 100 organoids and passaging at a 1 in 4 split ratio, organoid count increases an average of 4.2-fold per passage.
6 STEMCELL Technologies Inc. IntestiCult™ Organoid Why Use IntestiCult™ Organoid Growth Medium (Human) Growth Medium (Human)? Cell Culture Medium for ROBUST. Reliable and efficient expansion of intestinal stem cells across donor samples. Establishment and Maintenance RELEVANT. Enables generation of intestinal organoid of Human Intestinal Organoids cultures that can be used to model the adult intestinal epithelium. IntestiCult™ Organoid Growth Medium (Human) is a complete cell culture medium for efficient establishment and long-term COMPLETE. Two-component medium system does not maintenance of human intestinal epithelial organoids derived from require additional growth factors to support organoid primary intestinal or colonic crypts or previously frozen colonic growth. organoids. CONSISTENT. Quality-controlled medium generates consistent results between experiments and between laboratories.
A B DAPI EPCAM
Ki67 MERGE
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Figure 3. Human Colonic Organoids Grown in IntestiCult™ Organoid Growth Medium (Human)
(A) Colonic organoids grown in IntestiCult™ Organoid Growth Medium (Human) and imaged on day 7 of passage 0. (B) Immunofluorescence of intestinal organoids showing DAPI (blue), EPCAM (red), Ki67 (green), and the merged image.
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106 Cumulative Total Cell Yield Cumulative Total 105
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Figure 4. Intestinal Organoids Can be Maintained in Long-Term Figure 5. Forskolin-Induced Swelling of Intestinal Organoids
Culture Through Passaging Organoids cultured in IntestiCult™ Organoid Growth Medium (Human) were Organoids cultured in IntestiCult™ Organoid Growth Medium (Human) show treated with (A) 5 µM Forskolin or (B) DMSO. Organoid area was measured at efficient growth throughout passaging. Cultures were split with an average split 0 minutes and 120 minutes. Forskolin-treated organoids increased in size by ratio of 1 in 6 at each passage. Error bars represent standard error. 33.5 ± 3.8% in size compared to a 7.5 ± 0.8% increase for control organoids (n=3). Representative images are shown.
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IntestiCult™ Organoid Why Use IntestiCult™ Organoid Differentiation Medium Differentiation Medium? (Human) RELEVANT. Enables generation of intestinal organoid cultures with physiological proportions of differentiated Complete Culture Medium for the and stem cell populations. Differentiation of Intestinal Organoids. FLEXIBLE. Allows easy access to the apical surface by transitioning intestinal organoids into monolayer and ALI IntestiCult™ Organoid Differentiation Medium (Human; ODM) is a cultures. complete culture medium that supports the further differentiation REPRODUCIBLE. Generates consistent results across of intestinal organoids in three dimensions (3D), or in 2D as passages and replicates. submerged monolayers or air-liquid interface (ALI) cultures. These cultures can be initiated from intestinal organoids derived from human intestinal crypts, or passaged organoids that have been cultured with IntestiCult™ Organoid Growth Medium (Human; OGM; Catalog #06010).
A B Figure 6. Differentiated Human Intestinal Organoids Display a Budded Morphology
(A) Organoids grown in IntestiCult™ OGM are primarily cystic. (B) When switched to IntestiCult™ ODM, organoids develop a thickened epithelium with a pronounced, budded morphology indicative of a more differentiated state. Organoids were imaged on day 5 of expansion or differentiation respectively.
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A B Figure 7. Intestinal Organoids Contain a Higher Proportion of Mature Cell Types Following Differentiation in IntestiCult™ ODM
(A, B) Organoids grown in IntestiCult™ OGM are enriched for Ki-67+ proliferative cells (A), while containing few differentiated cell types such as goblet cells (MUC2, MUC2 CHGA A), enterocytes (KRT20, B), and enteroendocrine cells (CHGA, B). (C, D) When Ki-67 KRT20 + E-cadherin E-cadherin switched to IntestiCult™ ODM, organoids contain a small number of Ki-67 DAPI 100 µm DAPI 100 µm proliferative cells (C, arrows), with more physiological proportions of goblet cells (MUC2, C), enterocytes (KRT20, D), and chromogranin A- (CHGA-)positive enteroendocrine cells (D, arrow). C D
MUC2 CHGA Ki-67 KRT20 E-cadherin E-cadherin DAPI 100 µm DAPI 100 µm
8 STEMCELL Technologies Inc. A B C 1.5 Lgr5 1.5 Axin2 600 KRT20
1.0 1.0 400 RQ RQ RQ
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0.0 0.0 0 r r r d e er d er d er y y ye y ye noi a noi a a noi a a olay ol ol nol ga n ga n Orga on MUC2 Or CHG-A onol Or onol M Mo Mo Mo I IM IM D 40000 GM E 30000 GM M IntestiCult™GM OGMM Organoid O DM AL O D AL O D AL 30000 O ™ O ™ O ™ t ™ M t ™ M lt ™ DM 20000l t D l t D 20000 lt O l O l O ™ IntestiCult™ ODM™ Monolayer tiCu ™ tiCu t tiCu t s 10000 tiCu lt s tiCu l s tiCu l e s e 10000 s e s 120 e nt e nt e Int tiCu I 6000 nt tiCu I IntestiCult™nt ODMtiCu ALI Monolayer Int s I s I s 100 e e e 80 nt 4000 Int Int
RQ I RQ 60 2000 40 2.0 2.0 1.5 1.5 1.0 1.0 0.5 0.5 0.0 0.0 d r r er d er y ye y ye noi a a noi a a ol ol F ga n G ga n Or onol Or onol Mo Mo IM IM GM M GM M O D AL O D AL ™ O M MUC2™ O t ™ t ™ M l lt D l t D O E-cadherin l O ™ tiCu t tiCu ™ s tiCu l Ds API tiCu lt e s e s nt e e I nt tiCu Int tiCu I s Int s e e Int Int
MUC2 E-cadherin DAPI 30 µm 30 µm
Figure 8. Differentiation of Intestinal Epithelium at the Air-Liquid Interface (ALI) Using IntestiCult™ ODM
(A – E) Growing organoid-derived monolayers as ALI cultures drives further differentiation of intestinal epithelial cultures as seen by changes in gene expression measured by RT-qPCR. Relative quantification (RQ) for each marker is shown relative to actB and TBP housekeeping genes and normalized with respect to undifferentiated organoids grown in IntestiCult OGM (Human). Progenitor markers (A) Lgr5 and (B) Axin2 are significantly reduced in both submerged monolayers and ALI cultures, while markers of enterocytes (KRT20, C), goblet cells (MUC2, D), and enteroendocrine cells (CHGA, E) are significantly increased. Further reduction in Axin2 is seen in ALI monolayers with an increase in expression of KRT20, MUC2, and CHGA. (F, G) Comparing cross-sections of organoid monolayers grown in IntestiCult™ ODM as (F) submerged culture or (G) at the ALI shows further differentiation of the intestinal epithelium with an increased proportion of goblet cells and extracellular mucus (MUC2, green).
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A 2500 * *
0 220 IntestiCult™OGM DMSO OGM DMSO
2000 oT 200 IntestiCult™OGM Forskolin OGM Forskolin et 2 180 IntestiCult™ ODM DMSO m 1500 ODM DMSO
elativ 160 xc IntestiCult™ODM Forskolin ODM Forskolin 1000
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500 120 wellin 100 0 %S I 80 0 MIN 20 MIN 40 MIN 60 MIN co-2 AL Ca DM O Incubation Time t™ DM Monolayer l O tiCu ™ s lt e B IntestiCult™ ODM C Caco-2 Int tiCu s e Int IBMX/Forskolin CFTR inhibitor-172 40 IBMX/Forskolin CFTRinh172
100 30 2 2 Figure 9. Differentiated Organoid-Derived Monolayers and ALI 20 A/cm Cultures Display More Physiological Trans-Epithelial Electrical A/cm 50 10 µ µ
Resistance (TEER) than Caco-2 Cells 0
Differentiated organoid-derived monolayers grown as a submerged monolayer 0 -10 Time (IntestiCult™ ODM Monolayer), or at the ALI (IntestiCult™ ODM ALI), show higher Time TEER values as compared to Caco-2 cultures.Organoid-derived monolayers grown at the ALI show a loosening of tight junctions due to further differentiation of the D E IBMX/ForskolinIBMX/Forskolin CFTRCFTR Inhibitor-172 inhibitor-172 brush border, and thus lower TEER values are observed. * p < 0.0001 100 p < 0.001 100 p < 0.001 2) 2) 80 80
60 60 A/cm A/cm
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Figure 10. Differentiated Intestinal Organoids Provide a Suitable Model for Studying CFTR Response In Vitro
(A) Organoids differentiated further in IntestiCult™ ODM show a comparable degree of swelling when treated with forskolin as compared to organoids grown in IntestiCult™ OGM, demonstrating suitability for use in forskolin-induced swelling assays. (B – E) Ussing chamber analysis of submerged (B) organoid-derived monolayers and (C) Caco-2 cultures demonstrate increased sensitivity of organoid-derived monolayers to CFTR activation and inhibition by IBMX/Forskolin and CFTR Inhibitor-172 respectively. (D, E) Analysis of CFTR modulation by IBMX/ Forskolin and CFTR Inhibitor-172 show significantly greater (D) activation and (E) inhibition of CFTR activity in organoid-derived monolayers as compared to Caco-2 cultures (p < 0.001 for both).
10 STEMCELL Technologies Inc. STEMdiff™ Intestinal Why Use STEMdiff™ Intestinal Organoid Kit Organoid Kit? Cell Culture Medium Kit for RELEVANT. Enables generation of small intestinal organoid cultures that model the developing intestinal epithelium Differentiation of PSC-Derived and associated mesenchyme. Human Intestinal Organoids ROBUST. Supports efficient differentiation of human ES and iPS cell lines to intestinal organoids. The STEMdiff™ Intestinal Organoid Kit supports efficient establishment of PSC-derived small intestinal organoid cultures CONVENIENT. Intestinal organoids can be maintained from embryonic stem (ES) cells or induced pluripotent stem (iPS) long‑term through passaging or cryopreserved for cells within 30 days. This serum-free medium kit is based on the experimental flexibility. 3 formulation published by Spence et al. and has been optimized SERUM-FREE. Optimized formulation for low experimental to increase efficiency and reproducibility of organoid formation variability. and expansion.
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Seed Initiate Definitive Mid-/ Small Intestinal Human PSCs Differentiation Endoderm Hindgut Organoids
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Figure 11. Generation of Human Intestinal Organoid Cultures Using the STEMdiff™ Intestinal Organoid Kit
(A) Human PSC cultures progress through a three-stage differentiation process to generate human intestinal organoids. By day 3 of the protocol, cultures exhibit characteristics typical of definitive endoderm and mid-/hindgut differentiation is initiated. During mid-/hindgut differentiation (days 5 - 9), cells form mid-/hindgut spheroids that are released from the cell monolayer into the culture medium. These spheroids are collected and embedded in extracellular matrix. (B) Embedded mid-/hindgut spheroids cultured in STEMdiff™ Intestinal Organoid Growth Medium mature into intestinal organoids (days in parentheses indicate days post-embedding in a given passage). Once established, intestinal organoids can be maintained and expanded in culture by passaging every 7 - 10 days. After multiple passages, the organoids generally exhibit less sinking within the matrix dome and a lower proportion of mesenchymal cells.
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A B C
12001200 6000 H9 120 10001000 5000 H7 100100 WLS-1C + s
+ 800800 4000 STiPS-M001 8080
OX17 600600 3000 6060 / SOX17 pheroid /S + +
#S 400400 2000 4040 Cell Yield (x1000) Cell Yield OXA2
2020 # Spheroids per Well 200200 1000 %F %FOXA2 00 00 0 C ) ) C 09) 07) O1 -1 WA WA LS-1 MO WA09 WA07 LS 9 ( 7 ( W W S-MOO1 H H WLS-1C 9 ( 7 ( WLS-1C TiPS- H H P Spheroids Passage 1 Passage 2 Passage 3 H7 (WA07) S H7 (WA07) Ti H9 (WA09) STiPS-M001 H9 (WA09) SSTiPS-M001 Human Intestinal Organoids
Figure 12. STEMdiff™ Intestinal Organoid Kit Supports Robust Differentiation and Expansion Across ESC and iPSC Lines
STEMdiff™ Intestinal Organoid Kit enables high-efficiency generation of intestinal organoids from both ES cells (H9, H7) and iPS cells (WLS-1C, STiPS-M001). (A) Organoids initiated from a variety of cell lines show efficient induction of definitive endoderm, measured by co-expression of FOXA2 and SOX17 on day 3 of differentiation. (B) Both ES and iPS cell‑derived cultures demonstrate efficient spheroid formation upon mid-/hindgut induction. The total number of spheroids obtained per well in a given differentiation is shown. (C) Organoids cultured from either ES or iPS cells can be expanded and maintained over multiple passages. Shown is the total cell yield per passage. Organoids were passaged every 7 - 10 days with a split ratio between 1 in 2 and 1 in 4. Data points represent the mean of 3 biological replicates. Error bars throughout represent standard deviation of the mean.
Figure 13. Characteristics of Intestinal Organoids Cultured Using A B STEMdiff™ Intestinal Organoid Growth Medium
Intestinal organoids were stained and examined for marker expression by immunocytochemistry. (A,B) Intestinal organoids express markers of intestinal progenitor cells including CDX2 and the intestinal crypt marker SOX9. The organoids incorporate a DAPI polarized epithelium, visualized by the localization of EPCAM to the exterior (basolateral) CDX2 DAPI MUC2 CHGA surface of the organoids (A), and express markers typical of mature cell types including EPCAM 100 µm SOX9 100 µm MUC2 (goblet cells, A) and CHGA (enteroendocrine cells, B). (C,D) Observation of desmin (C) and vimentin (D) in intestinal organoids demonstrates incorporation of mesenchymal C D cells in the organoid cultures, while KRT20 (C) and Ki67 (D) are markers of differentiated intestinal cells and actively dividing progenitor cells, respectively. Images are digital cross- sections of whole-mount imaging of intact organoids.
DAPI DAPI KRT20 Ki67 Desmin Vimentin EPCAM 100 µm EPCAM 100 µm
12 STEMCELL Technologies Inc. Hepatic Organoids
The development of hepatic organoid culture techniques has provided the hepatic research field with a convenient method for sustained maintenance of liver cells in vitro.4,5 Hepatic progenitor organoids are cultured by isolating and expanding liver stem and progenitor cells, which are postulated to reside in hepatic ducts. Culture of hepatic progenitor cells as organoids produces spherical organoids consisting primarily of progenitor cells that can be further differentiated to either hepatocytes or cholangiocytes.6
HepatiCult™ Organoid Why Use HepatiCult™ Organoid Growth Medium (Mouse) Growth Medium (Mouse)? Cell Culture Medium for Establishment CONVENIENT. In vitro system for generating organoids within a week. and Maintenance of Mouse Hepatic STEP-BY-STEP PROTOCOL. No injury models, hand- Organoids picking of ducts, or cell sorting required.
HepatiCult™ Organoid Growth Medium (Mouse) is a serum-free SIMPLE, TWO-COMPONENT FORMAT. Serum-free medium that allows for rapid generation of hepatic progenitor medium formulation. organoids from mouse liver tissue. Organoid culture may be initiated FLEXIBLE PROTOCOL. Organoids be can grown from duct from hepatic ducts, duct fragments, single cells, or cryopreserved fragments or single cells and cultured in matrix domes or ® ® organoids and cultured in Corning Matrigel domes or as a dilute suspension. Matrigel® suspension.
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Figure 14. Mouse Hepatic Progenitor Organoids Can Be Initiated from a Variety of Starting Materials
HepatiCult™ Organoid Growth Medium (Mouse) enables the initiation of hepatic progenitor organoids from (A) duct fragments, (B) single cells or (C) cryopreserved organoids. All organoids were grown in Matrigel® domes and imaged on day 7 of primary culture or the first passage post thaw (cryopreserved organoids).
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Figure 15. Hepatic Organoids Can Be Grown in Matrigel® Domes or in a Dilute Matrigel® Suspension
Hepatic progenitor organoids cultured from freshly isolated mouse hepatic duct fragments in HepatiCult™ Organoid Growth Medium (Mouse) can be plated in (A) Matrigel® domes or (B) a dilute Matrigel® suspension. Organoids grown in either culture condition are ready for passage within 4-7 days.
Dome-Expanded Suspension-Expanded 10251025 Hepatic Organoids Hepatic Organoids ield Proliferation Mki67 ly Axin2 201020 Cd24a 10 cel Cd44 Prom1 15 15 Sox9 total 10 10 Wnt Pathway Lgr5 Epcam Epithelial Tjp1 10101010 Acox2 Cumulative Total Cell Yield Cumulative Total Foxa2 Hnf1α cumulative 105 105 Hnf4α p0 p1 p2 p3 p4 p5 p6 p7 p8 p9 p10
Hepatic Pparg p0 p1 p2 p3 p4 p5 p6 p7 p8 p9 p10 Progenitor Tbx3 Passage Number Ttr Hnf1β Figure 17. Expansion of Organoids Grown in HepatiCult™ Ductal Krt7 Progenitor Organoid Growth Medium (Mouse) Krt19 Afp Organoids cultured with HepatiCult™ Organoid Growth Medium (Mouse) show Ldlr -2 Abcc4 efficient growth over multiple passages.Cultures were split with an average split )
2 Alb ratio of 1:25 at each passage. Asgr1 Cyp3a11 Hepatic 8 Fah Glul Ppara Serpina1a
Gene Expression(rlog Gene Ggt1 16 Notch2 Onecut1 Ductal Sstr2 Pou5f1 Pluripotency Sox2
Figure 16. Analysis of Organoid Gene Expression
Analysis of marker expression by RNA-seq shows organoids grown in HepatiCult™ Organoid Growth Medium (Mouse) in either Matrigel® domes or in a dilute Matrigel® suspension express markers associated with hepatic stem and progenitor cells. The organoids also express low levels of genes associated with mature hepatic cell types, including cholangiocytes and hepatocytes. Columns represent biological replicates at passages ranging from passage 1 - 40. 14 STEMCELL Technologies Inc. A B A Hnf4α B Alb n α l . RQ RQ