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STEMdiff™肠道类器官生长培养基

用于长期维持 hPSC 衍生的人类肠道类器官的无血清培养基
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产品号 #05145_C

用于长期维持 hPSC 衍生的人类肠道类器官的无血清培养基

产品组分包括

  • STEMdiff™ Intestinal Organoid Basal Medium, 100 mL
  • STEMdiff™ Intestinal Organoid Supplement, 2 mL
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应用领域
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总览

长期维持人类肠道类器官作为研究肠道研究、疾病建模、药物发现和化合物筛选的模型系统。使用无血清STEMdiff™肠道类器官生长培养基（OGM）对人类多能干细胞（hPSC）衍生的肠道类器官进行扩展、一致的培养。

根据Spence et al. （Nature 2011）发表的配方，STEMdiff™Intestinal OGM经过优化，可在hPSC系间高效、可重复地产生和扩增类器官。使用STEMdiff™肠OGM从造血干细胞分化成熟的中/后肠球体STEMdiff™肠道类器官试剂盒． 球体会成熟为肠道类器官，你可以通过传代冷冻保存或长期保存。在STEMdiff™肠道OGM中培养的肠道类器官显示出模拟发育中的肠上皮和相关间质的细胞组成和组织，使其成为与发育中的肠道直接相关的方便的模型系统。

细胞类型
内胚层，PSC衍生，肠道细胞
 
种属
人
 
应用
细胞培养，分化，类器官培养
 
品牌
STEMdiff
 
研究领域
疾病建模，药物发现和毒理检测，上皮细胞研究
 
制剂类别
无血清
 

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Data Figures

Figure 1. STEMdiff™ Intestinal Organoid Kit Enables the Growth of Intestinal Organoids from PSCs

STEMdiff™ Intestinal Organoid Kit facilitates directed differentiation of PSCs to form human small intestinal organoids. The organoids exhibit a polarized epithelial monolayer surrounding a hollow lumen and are associated with a mesenchymal cell population. Pictured are passage 3 human intestinal organoids generated using this kit.

Figure 2. 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.

Figure 3. 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 ESCs (H9, H7) and iPSCs (WLS-1C, STiPS-MOO1). (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 ESCand iPSC-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 ESCs or iPSCs 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 4. Characteristics of Mid-/Hindgut Spheroids Generated with STEMdiff™ Intestinal Organoid Kit

(A) Cultures differentiated using STEMdiff™ Intestinal Organoid Kit exhibit the expected markers during definitive endoderm and mid-/hindgut specification. During the protocol, gene expression patterns shift from pluripotency markers (day 0) to definitive endoderm markers by day 3 and those of the mid-/hindgut epithelium by day 9. Mid-/hindgut cultures (day 9) also express markers of the associated mesenchyme. Marker levels were assessed by RT-qPCR and normalized to expression levels for undifferentiatied H9 cells. (B) Mid-/hindgut spheroids (day 9) express markers of the intestinal epithelium (CDX2, E-cadherin, EPCAM). (C) Mid-/hindgut spheroids (day 9) also incorporate components of the associated mesenchyme (vimentin).

Figure 5. Intestinal Organoids Cultured In STEMdiff™ Intestinal Organoid Growth Medium Exhibit Features of the Intestinal Epithelium

(A) Differentiated PSC-derived intestinal organoids express markers of the intestinal epithelium and the associated mesenchyme. Marker levels were assessed by RT-qPCR and normalized to expression levels for undifferentiated H9 cells. (B,C) Intestinal organoids express markers of intestinal progenitor cells including CDX2 and the intestinal crypt marker SOX9. The organoids are composed of a polarized epithelium, visualized by the localization of EPCAM to the exterior (basolateral) surface of the organoids (B), and express markers typical of mature cell types including MUC2 (B: goblet cells) and CHGA (C: enteroendocrine cells). (D,E) Observation of desmin (D) and vimentin (E) in intestinal organoids demonstrates incorporation of mesenchymal cells in the organoid cultures, while KRT20 (D) and Ki67 (E) are markers of differentiated intestinal cells and putative intestinal stem cells, respectively. Images are digital cross-sections of whole-mount immunofluorescence-stained intestinal organoids at P28 (Day 7).

Immunocytochemistry image of an intestinal organoid cultured in mTeSR™ Plus and directed to intestinal organoids using the STEMdiff™ Intestinal Organoid Kit.

Figure 6. Generation of Intestinal Organoids from hPSCs Maintained in mTeSR™ Plus

Human ES (H9) cells were cultured with mTeSR™ Plus and directed to intestinal organoids using the STEMdiff™ Intestinal Organoid Kit. Image shows markers of the intestinal epithelium EpCAM (green) and CDX2 (red), and intestinal mesenchyme marker vimentin (white). Nuclei are counterstained with DAPI (blue).

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
STEMdiff™ Intestinal Organoid Growth Medium
Catalog #
05145
Lot #
All
Language
English
Document Type
Technical Manual
Product Name
STEMdiff™ Intestinal Organoid Growth Medium
Catalog #
05145
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
STEMdiff™ Intestinal Organoid Growth Medium
Catalog #
05145
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
STEMdiff™ Intestinal Organoid Growth Medium
Catalog #
05145
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
STEMdiff™ Intestinal Organoid Growth Medium
Catalog #
05145
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
STEMdiff™ Intestinal Organoid Growth Medium
Catalog #
05145
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages
Workflow Stages for Organoids
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Resources and Publications

Educational Materials (40)

STEMdiff™ Intestinal Organoid Kit
Brochure
STEMdiff™ Intestinal Organoid Kit
Organoids
Brochure
Organoids
Intestinal Organoids
Brochure
Intestinal Organoids
CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
Protocol
CRISPR-Cas9 Genome Editing of Human Intestinal Organoids
How to Differentiate hiPSCs into Colonic Organoids Using STEMdiff™
Protocol
How to Differentiate hiPSCs into Colonic Organoids Using STEMdiff™
Barrier Permeability Assay Using Human Intestinal Organoid-Derived Monolayers
Protocol
Barrier Permeability Assay Using Human Intestinal Organoid-Derived Monolayers
Performing Immunocytochemical Staining of Epithelial Organoids
Protocol
Performing Immunocytochemical Staining of Epithelial Organoids
How to Prepare Single Cells from PSC-Derived Epithelial Cultures for Flow Cytometry Analysis
Protocol
How to Prepare Single Cells from PSC-Derived Epithelial Cultures for Flow Cytometry Analysis
How to Process Epithelial Organoids and Organoid-Derived Epithelial Monolayers for RNA Isolation
Protocol
How to Process Epithelial Organoids and Organoid-Derived Epithelial Monolayers for RNA Isolation
How to Perform Immunocytochemistry (ICC) Staining of Epithelial Cells Cultured as Monolayers or at the Air-Liquid Interface
Protocol
How to Perform Immunocytochemistry (ICC) Staining of Epithelial Cells Cultured as Monolayers or at t...
Derivation and Applications of Human Pluripotent Stem Cells
Wallchart
Derivation and Applications of Human Pluripotent Stem Cells
SnapShot: Growing Organoids from Stem Cells
Wallchart
SnapShot: Growing Organoids from Stem Cells
How to Count Intestinal Crypts for Mouse Organoid Cultures
1:09
Video
How to Count Intestinal Crypts for Mouse Organoid Cultures
Supporting Science Globally: A look into how scientists in Singapore are using organoids
4:46
Video
Supporting Science Globally: A look into how scientists in Singapore are using organoids
Differentiation of High-Quality hPSCs Using STEMdiff™ and TeSR™-AOF
12:44
Webinar
Differentiation of High-Quality hPSCs Using STEMdiff™ and TeSR™-AOF
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
56:31
Webinar
Modeling Human Gastrointestinal Development and Disease Using Pluripotent Stem Cells
GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
24:56
Webinar
GI Tract Organoids: Using Advanced Tissue Models to Interrogate Absorption and Regulation
Spotlight on Organoids: Expert Panel
Webinar
Spotlight on Organoids: Expert Panel
Nature Research Round Table: The Promise of Organoid Medicine
17:17
Webinar
Nature Research Round Table: The Promise of Organoid Medicine
Nature Research Round Table: Self-Organization and Symmetry Breaking in Intestinal Organoid Development
11:52
Webinar
Nature Research Round Table: Self-Organization and Symmetry Breaking in Intestinal Organoid Developm...
Dissecting the Innate Immune Response in Human Intestinal Organoid Models
55:56
Webinar
Dissecting the Innate Immune Response in Human Intestinal Organoid Models
Patient-Derived Organoids for Drug Screening and Development
50:25
Webinar
Patient-Derived Organoids for Drug Screening and Development
Nature Research Round Table: Organoids as Models for Human Disease
15:10
Webinar
Nature Research Round Table: Organoids as Models for Human Disease
From Stem Cells to Organoids: Modeling the Gastrointestinal Tract
1:03:40
Webinar
From Stem Cells to Organoids: Modeling the Gastrointestinal Tract
Organoid Expert Panel
52:35
Webinar
Organoid Expert Panel
Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
17:20
Webinar
Nature Research Round Table: Organoids as an Enabling Technology for Precision Cancer Medicine
Nature Research Round Table: Organoid Modeling of Stem Cells and Disease Microenvironments
4:47
Webinar
Nature Research Round Table: Organoid Modeling of Stem Cells and Disease Microenvironments
Nature Research Round Table: Modeling Intestinal Development In Vivo and In Vitro
10:17
Webinar
Nature Research Round Table: Modeling Intestinal Development In Vivo and In Vitro
Advances in GI Organoid Applications: Drug Discovery and Immunotherapy
47:49
Webinar
Advances in GI Organoid Applications: Drug Discovery and Immunotherapy
Challenges in Translating iPSC Technology
41:47
Webinar
Challenges in Translating iPSC Technology
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
1:10:09
Webinar
HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
15:15
Webinar
Nature Research Round Table: Modeling Congenital Pediatric Diarrhea in Intestinal Enteroids
Engineering Organoids Using ECM from Decellularized Tissue
36:39
Webinar
Engineering Organoids Using ECM from Decellularized Tissue
Nature Research Round Table: Progress and Challenges in Organoid Models of Human Brain Development
9:36
Webinar
Nature Research Round Table: Progress and Challenges in Organoid Models of Human Brain Development
Highly Efficient Differentiation of Human Pluripotent Stem Cells into Long-term Expandable “Mini-gut” Organoids
Scientific Poster
Highly Efficient Differentiation of Human Pluripotent Stem Cells into Long-term Expandable “Mini-g...
Caroline Lindemans, MD, PhD
Interview
Caroline Lindemans, MD, PhD
Heather McCauley, PhD
Interview
Heather McCauley, PhD
Maartje van der Heijden
Interview
Maartje van der Heijden
Jason Spence
Interview
Jason Spence
Tamara Zietek, PhD
Interview
Tamara Zietek, PhD

更多信息

更多信息
种属 Human
配方类别 Serum-Free
Legal Statement: STEMdiff™ Intestinal Organoid Kit was developed under license to intellectual property owned by the Cincinnati Children’s Hospital. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
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