For ex vivo clonogenicity assay, crypts were isolated from the small intestine of tamoxifen treated mice as described above. Crypts were counted and seeded in matrigel as described. The formation of organoids was assessed upon 2–3 days of culture in standard organoid growth medium (Stemcell Technologies). For the secondary organoid culture, primary organoids were mechanically dissociated, washed twice with PBS and embedded in matrigel. Growth of secondary organoids in standard organoid growth medium (Stemcell Technologies) was assessed after 24 h of culture.
Li S., Lu C.W., Diem E.C., Li W., Guderian M., Lindenberg M., Kruse F., Buettner M., Floess S., Winny M.R., Geffers R., Richnow H.H., Abraham W.R., Grassl G.A, & Lochner M. (2022). Acetyl-CoA-Carboxylase 1-mediated de novo fatty acid synthesis sustains Lgr5+ intestinal stem cell function. Nature Communications, 13, 3998.
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