Healthy human colonoids (from FARMM study) were generated and propagated in Matrigel (Corning, 356231) on 24-well polystyrene plates as previously described (48) with full growth medium comprising basal medium (advanced DMEM supplemented with 10% FBS, 1× penicillin/streptomycin, 1× GlutaMAX, 10 mM HEPES, 1× N2 supplement, 1× B27 supplement, all from Invitrogen), 1 mM N-acetyl-cysteine, 10 nM leu-gastrin (all from MilliporeSigma) and WRN cell conditioned medium (WRN cells from ATCC; basal medium and WRN conditioned medium mixed at 1:1 ratio), and supplements (500 nM A83-01 [Tocris], 10 μM SB-20210, 10 mM nicotinamide, 50 ng/mL human EGF [all from MilliporeSigma], and 2 μL/mL 1× Primocin [InvivoGen]). For the first 48 hours of colonoid culture, growth medium was supplemented with 10 μM Y27632, 2.5 μM CHIR99021 (both from MilliporeSigma), and 2.5 μM thiazovivin (Reprocell). Media was replaced every 48 hours.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.