Abstract:
:An important practical limitation of the three-dimensional geometry of stem-cell derived intestinal organoids is that it prevents easy access to the apical epithelium for testing food components, microorganisms, bioactive and toxic compounds. To this end, we here report on a new robust method for generating confluent intestinal cell monolayers from single-cell suspensions of enzymatically-dissociated porcine organoids using modified culture conditions. With this method, cell seeding densities can be standardised, overcoming problems with methods based on mechanical dissociation of organoids. Confluent monolayers formed tight junctions with high transepithelial electrical resistance in three days and could be used in experiments for up to two weeks. Multilineage differentiation of ileal stem cells was demonstrated by immunohistochemistry and RT-qPCR of cell-specific transcripts, also unequivocally confirming the controversial existence of Paneth-like cells in the porcine small intestine. The method described here is useful to standardize primary epithelial monolayer formation from intestinal organoids and allows rapid and robust studies of intestinal physiology.
journal_name
Stem Cell Resjournal_title
Stem cell researchauthors
van der Hee B,Loonen LMP,Taverne N,Taverne-Thiele JJ,Smidt H,Wells JMdoi
10.1016/j.scr.2018.02.013subject
Has Abstractpub_date
2018-04-01 00:00:00pages
165-171eissn
1873-5061issn
1876-7753pii
S1873-5061(18)30057-6journal_volume
28pub_type
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journal_title:Stem cell research
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