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.
SCM104
3dGRO® R-Spondin-1 Conditioned Media Supplement
Alternative stem cell media supplement to purified recombinant R-Spondin-1 protein for organoid cultures.
Synonym(s):
3dGRO Media Supplement, Conditioned Media Supplement, R-Spondin-1 Supplement
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Disclaimer
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.
General description
3D organoid culture systems are increasingly employed as powerful tools for the study of human diseases. R-Spondin proteins are a family of cysteine-rich, thrombospondin type I repeat containing proteins that activate both the WNT/β-catenin and WNT/PCP pathways. R-spondin-1 (RSPO1) is one of the most extensively used niche factors for culturing 3D organoids and has been used to establish organoids from the stomach, small intestine, colon, pancreas and liver from both mouse and human sources1,2,3 along with intestinal organoids from other large and small animals4. However, due to the high concentration requirement in organoid media the use of the recombinant R-spondin-1 becomes prohibitively expensive. The 3dGRO® R-Spondin-1 Conditioned Media Supplement is a potent and inexpensive alternative to purified recombinant RSPO1 protein for organoid cultures and is optimized for the highest level of RSPO-1 expression to ensure consistency and performance.
Source:
The 3dGRO® R-Spondin-1 Conditioned Media Supplement is generated from a 293T cell transfectant stably secreting mouse R-spondin-1 tagged with an influenza hemagglutinin (HA) epitope at the N-terminus and a murine IgG2a Fc fragment at the C-terminus. The conditioned medium does not contain serum.
Source:
The 3dGRO® R-Spondin-1 Conditioned Media Supplement is generated from a 293T cell transfectant stably secreting mouse R-spondin-1 tagged with an influenza hemagglutinin (HA) epitope at the N-terminus and a murine IgG2a Fc fragment at the C-terminus. The conditioned medium does not contain serum.
Preparation Note
RSPO-1 Conditioned Medium should be stored at -20°C. When ready to use, thaw the supplement overnight at 2-8°C in the dark. Thawed RSPO-1 Conditioned Medium is good for two weeks when stored at 2-8°C.
Legal Information
3dGRO is a registered trademark of Merck KGaA, Darmstadt, Germany
Storage Class
12 - Non Combustible Liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
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How Do you Extract RNA/DNA from Organoids in Cultured in Matrigel?
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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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Dear official, Dear official What is the concentration of the R-Spondin-1 Conditioned Media Supplement (#SCM104) product and I could not find the datasheet on the site, can you give me information?
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This product is a 2X medium supplement. The material is a conditioned medium from 293T cells that have been transfected to produce R-spondin-1. The base formulation is considered proprietary. Please see the link below to review the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/404/988/scm104ds.pdfHelpful?
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