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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.
SCM105
3dGRO® L-WRN Conditioned Media Supplement
3dGRO® L-WRN Conditioned Media Supplement
1 of 1 reviewers received a sample product or took part in a promotion
Synonym(s):
Conditioned Media
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About This Item
UNSPSC Code:
12352207
NACRES:
NA.71
Pricing and availability is not currently available.
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form
liquid
manufacturer/tradename
Millipore
technique(s)
cell based assay: suitable
color
clear red
shipped in
ambient
storage temp.
−20°C
Application
Appearance (color): clear, red liquid
Endotoxin: <2 EU/mL
Sterility Tested: No Growth/Pass
Media are tested for function in supporting growth of human intestinal organoids
Media are tested for levels of growth factors by ELISA
Endotoxin: <2 EU/mL
Sterility Tested: No Growth/Pass
Media are tested for function in supporting growth of human intestinal organoids
Media are tested for levels of growth factors by ELISA
The 3dGRO® L-WRN Conditioned Media Supplement is a formulation based on DMEM/F-12 PLUS medium containing the 3 growth factors Wnt3a, R-spondin-3, and Noggin. These factors act upon the Wnt signaling pathway, which plays a critical role in sustaining the proliferation of LGR5+ crypt stem cells within organoids. Wnt activation is thus essential for in vitro growth of human gut organoids. L-WRN conditioned medium is widely used as a component of the culture medium for supporting the growth of human intestinal organoids including the 3dGRO® gastrointestinal organoid biobank (SCC310-SCC367). Applications for L-WRN conditioned medium include but are not limited to the expansion of human intestinal gut organoids and other epithelial organoid cell types.
Source
The 3dGRO® L-WRN Conditioned Media Supplement is derived from the Mus musculus L-M(TK-) cell line secreting mouse Wnt3a, R-spondin-3 and Noggin. L-WRN Conditioned Medium may contain trace amounts of mouse DNA.
Source
The 3dGRO® L-WRN Conditioned Media Supplement is derived from the Mus musculus L-M(TK-) cell line secreting mouse Wnt3a, R-spondin-3 and Noggin. L-WRN Conditioned Medium may contain trace amounts of mouse DNA.
Features and Benefits
L-WRN conditioned media supplement containing the 3 growth factors Wnt3a (W), R-spondin-3 (R), and Noggin (N) for organoid cultures.
Storage and Stability
The 3dGRO® L-WRN Conditioned Media Supplement should be stored at -20°C. When ready to use, thaw the L-WRN Conditioned Medium overnight at 2-8°C in the dark. Thawed medium is good for two weeks when stored at 2-8°C. If media will not be used within two weeks, aliquot unused media and store at -20°C. After thawing refrozen aliquots, do not re-freeze media.
Legal Information
3dGRO is a registered trademark of Merck KGaA, Darmstadt, Germany
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.
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
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 are organoids cultured?
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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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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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Is there a version of this product without Wnt3a, i.e., only with Noggin and R-spondin-3 ? Can you buy Noggin conditioned media separately?
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There are no such conditioning medias available as you describe. The only other organoid conditioned media is product 3dGRO™ R-Spondin-1 Conditioned Media Supplement, this is product SCM104: https://www.sigmaaldrich.com/US/en/product/mm/scm104.
We recommend contacting our Sales team who can assist you with further possible options of a custom product. Please navigate to the link https://www.sigmaaldrich.com/support/customer-support, click on "Quotes" under the Orders Section, and provide the required information. Thank you.
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