Mouse colon stem cells were cultured using IntestiCult Organoid Growth Media according to the manufacturer’s instructions (06005, STEMCELL Technologies). The whole colon was removed from untreated WT and Nlrc3−/− mice and rinsed with cold PBS. The colon was cut into 2 mm segments and washed 20 times with cold PBS. Colonic segments were incubated in Gentle Cell Dissociation Reagent (07174, STEMCELL Technologies), rotated at 350 g for 15 min at room temperature, followed by resuspension in 0.1% BSA (A6003, Sigma) in PBS. Dissociated colonic crypts were filtered through 70-µm strainers. Dissociated colonic crypts were resuspended in DMEM/F12 medium with 15 mM HEPES (36254, STEMCELL Technologies), counted and resuspended in IntestiCult Organoid Growth Media and Matrigel (356230, Corning) in a 1:1 ratio. Cells were plated in 24-well culture plates (3738, Corning). IntestiCult Organoid Growth Media were added to the cell culture plates to immerse the matrix composed of IntestiCult Organoid Growth Media and Matrigel. For inhibition studies, 50 µM of LY294002 (440202, Millipore) or 10 µg/ml rapamycin (553210, Sigma) was added to the IntestiCult Organoid Growth Media.
Intesticult organoid growth media
Manufactured by Corning
Intesticult Organoid Growth Media is a specialized culture medium designed to support the growth and maintenance of intestinal organoids. It provides the necessary nutrients and growth factors to promote the formation and differentiation of these three-dimensional cell culture models derived from intestinal stem cells.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by Corning (2 mentions)
Ly294002, by Merck Group (2 mentions)
Rapamycin, by Merck Group (2 mentions)
Gentle cell dissociation reagent (gcdr), by STEMCELL (2 mentions)
Intesticult organoid growth media, by STEMCELL (2 mentions)
Hepes, by STEMCELL (2 mentions)
A6003, by Merck Group (2 mentions)
2 protocols using intesticult organoid growth media
1
Culturing Mouse Colon Stem Cells
2
Culturing Mouse Colon Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mouse colon stem cells were cultured using IntestiCult Organoid Growth Media according to the manufacturer’s instructions (06005, STEMCELL Technologies). The whole colon was removed from untreated WT and Nlrc3−/− mice and rinsed with cold PBS. The colon was cut into 2 mm segments and washed 20 times with cold PBS. Colonic segments were incubated in Gentle Cell Dissociation Reagent (07174, STEMCELL Technologies), rotated at 350 g for 15 min at room temperature, followed by resuspension in 0.1% BSA (A6003, Sigma) in PBS. Dissociated colonic crypts were filtered through 70-µm strainers. Dissociated colonic crypts were resuspended in DMEM/F12 medium with 15 mM HEPES (36254, STEMCELL Technologies), counted and resuspended in IntestiCult Organoid Growth Media and Matrigel (356230, Corning) in a 1:1 ratio. Cells were plated in 24-well culture plates (3738, Corning). IntestiCult Organoid Growth Media were added to the cell culture plates to immerse the matrix composed of IntestiCult Organoid Growth Media and Matrigel. For inhibition studies, 50 µM of LY294002 (440202, Millipore) or 10 µg/ml rapamycin (553210, Sigma) was added to the IntestiCult Organoid Growth Media.
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