Colon spheroids will form at the end of Day 12, which will be embedded in Matrigel® matrix using the 3dGRO™ Human Colon Organoid Expansion Media (SCM304) to generate mature colonic organoids. By day 10-12 in culture, colon organoids occupy 85-90% of the dome, and are ready to be passaged. For reference, below is the link to In Vitro Differentiation of Human iPS Cells Into Colon Organoids in Serum-Free Cell Culture Conditions:
SCC300
3dGRO® Human iPSC Derived Colon Organoids
Synonym(s):
Human Colon Organoids, Human Colonoids, Human Gut Organoids, Human Intestinal Organoids
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About This Item
UNSPSC Code:
41106514
NACRES:
NA.81
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form
liquid
technique(s)
cell culture | stem cell: suitable
application(s)
cell analysis
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| technique(s) cell culture | stem cell: suitable | technique(s) cell culture | mammalian: suitable, cell culture | stem cell: suitable | technique(s) cell culture | mammalian: suitable, cell culture | stem cell: suitable | technique(s) cell culture | mammalian: suitable, cell culture | stem cell: suitable |
| form liquid | form - | form - | form - |
| application(s) cell analysis | application(s) - | application(s) - | application(s) - |
General description
Human intestinal organoids are self-organizing, 3D structures that can be expanded in long-term culture and differentiate into intestinal epithelial cell types. Epithelial intestinal organoids, often referred to as enteroids or “mini-guts”, maintain the physiological characteristics of the gastrointestinal system and have been a useful cell culture tool to model intestinal development and disease including the study of colon cancer, celiac disease, inflammatory bowel diseases and host microbiome interactions. Organoids can be propagated from patient biopsies and from embryonic (ES) and induced pluripotent stem cells (iPS cells).
3dGRO® Human iPSC Derived Colon Organoids are derived from integration-free human iPS cells (Cat. No. SCC271) reprogrammed using the Simplicon RNA reprogramming technology (Cat. No. SCR550). Human iPS cells were differentiated using a multi-step process from definitive endoderm to posterior hind-gut endoderm and eventually into colon organoids (Figure 1.). The organoids express colon-specific markers including the posterior hindgut marker CDX2, a-carbonic anydrase II (CA-II), a-carbonic anhydrase IV (CA-IV), and goblet cell markers Mucin-2 and Mucin-5B. 3dGRO® human iPSC derived colon organoids can be propagated long-term by passaging every 10-12 days using 3dGRO® Human Colon Organoid Expansion Medium (Cat. No. SCM304).
3dGRO® Human iPSC Derived Colon Organoids are derived from integration-free human iPS cells (Cat. No. SCC271) reprogrammed using the Simplicon RNA reprogramming technology (Cat. No. SCR550). Human iPS cells were differentiated using a multi-step process from definitive endoderm to posterior hind-gut endoderm and eventually into colon organoids (Figure 1.). The organoids express colon-specific markers including the posterior hindgut marker CDX2, a-carbonic anydrase II (CA-II), a-carbonic anhydrase IV (CA-IV), and goblet cell markers Mucin-2 and Mucin-5B. 3dGRO® human iPSC derived colon organoids can be propagated long-term by passaging every 10-12 days using 3dGRO® Human Colon Organoid Expansion Medium (Cat. No. SCM304).
Application
Highly characterized cryopreserved human colon intestinal organoids derived from integration-free human induced pluripotent stem cells (iPSCs).
Research Category:
Research Category:
- Cell Culture
- Stem Cell Research
Quality
- Each vial contains ≥1500 organoids.
- Mycoplasma: Negative
- Genotyped by STR analysis: Pass
- Human species verified
- Organoids are negative for HPV-16, HPV-18, Hepatitis A, B, C, and HIV-1 & 2 viruses by PCR.
- Cell viability test: Pass
Storage and Stability
3dGRO® Human iPSC Derived Colon Organoids should be stored in liquid nitrogen.
Legal Information
3dGRO is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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How long before it is ready to use?
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How are organoids cultured?
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Once isolated, organoids are cultured in specialized media embedded in a rich extracellular matrix (ECM) basement membrane extract hydrogel such as Growth Factor Reduced Matrigel. Media is replenished every other day and cells are passaged once per week (7-12 days) before cells become too large or necrotic. Cells can be passaged in either small clump fragments or as single cells using mechanical dissociation/enzyme-free passaging reagents. Depending on organoid type and confluency, split ratios of roughly 1:3-1:4 can be used when passaging organoids. For sensitive organoid types, ROCKi (Y27632) (SCM075) can be added during passaging to help in promoting cell viability.
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How do you stain organoids with antibodies?
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Organoids can be stained with antibodies for immunocytochemical (ICC)/Immunohistochemical (IHC) analysis using either whole-mount or paraffin embedding/sectioning techniques. It is critical to use antibodies prequalified for organoid analysis.
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How Do you Extract RNA/DNA from Organoids in Cultured in Matrigel?
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Traditional DNA/RNA isolation kits can be used to prepare genomic material for sequencing (RNA-seq, scRNA-seq, WGS, CHIP-seq etc). For some applications with large organoids, tissue specific DNA/RNA extraction kits are recommended. TRI Reagent (T9424) can be added to the dissociated organoids before RNA extraction and can be stored at -80 °C until future use.
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