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.
SCM162
DMEM/F-12 PLUS Basal Medium
Advanced DMEMF12 basal media used in low-serum or serum-free media formulations including stem cell and 3D organoid cell cultures.
Synonym(s):
Advanced DMEM/F12, Organoid Basal Media, Organoid Basal Media, DMEMF12, DMEM F12, Dulbecco′s Modified Eagle Medium/Ham′s F-12, Serum-Free Basal Media
About This Item
Recommended Products
form
liquid
technique(s)
cell culture | stem cell: suitable
General description
• Culture of primary, stem cells and cell lines in applications that require reduced or serum-free medium.
• Optimization of transfections that require reduced or serum-free conditions.
• Production of conditioned medium in reduced serum or serum-free conditions.
• Optimized media for the culture and expansion of multiple 3D organoids and tissues systems.
DMEM/F12 PLUS is a proprietary formulation that contains human serum albumin (HSA), defined lipids, trace elements, chemicals and proteins essential for optimal cell growth in reduced serum conditions. The media contains high-glucose and does not contain L-glutamine or penicillin-streptomycin.
Application
Stem Cell Research
Quality
Osmolality: 290-330 mOsm
pH: 7.2-7.4
Sterility Tested: No Growth/Pass
Endotoxin: <2 EU/mL
Mycoplasma: Negative
Storage and Stability
Disclaimer
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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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Protocols
A rapid in vitro assay for CFTR function, the forskolin-induced swelling protocol uses human colon organoids, which can be derived from cystic fibrosis patient tissue.
How to stain organoids? A complete step-by-step protocol for immunofluorescent (IF) and immunocytochemical (ICC) staining of organoid cultures using antibodies
How to stain organoids? A complete step-by-step protocol for immunofluorescent (IF) and immunocytochemical (ICC) staining of organoid cultures using antibodies
How to stain organoids? A complete step-by-step protocol for immunofluorescent (IF) and immunocytochemical (ICC) staining of organoid cultures using antibodies
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How do you stain organoids with antibodies?
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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 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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