Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 165-171 |
| Journal | Stem Cell Research |
| Volume | 28 |
| DOIs | |
| Publication status | Published - 1 Apr 2018 |
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Keywords
- Epithelium
- Intestinal
- Monolayer
- Organoids
- Porcine
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Optimized procedures for generating an enhanced, near physiological 2D culture system from porcine intestinal organoids. / van der Hee, B.; Loonen, L.M.P.; Taverne, N.; Taverne-Thiele, J.J.; Smidt, H.; Wells, J.M.
In: Stem Cell Research, Vol. 28, 01.04.2018, p. 165-171.Research output: Contribution to journal › Article › Academic › peer-review
TY - JOUR
T1 - Optimized procedures for generating an enhanced, near physiological 2D culture system from porcine intestinal organoids
AU - van der Hee, B.
AU - Loonen, L.M.P.
AU - Taverne, N.
AU - Taverne-Thiele, J.J.
AU - Smidt, H.
AU - Wells, J.M.
PY - 2018/4/1
Y1 - 2018/4/1
N2 - 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.
AB - 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.
KW - Epithelium
KW - Intestinal
KW - Monolayer
KW - Organoids
KW - Porcine
U2 - 10.1016/j.scr.2018.02.013
DO - 10.1016/j.scr.2018.02.013
M3 - Article
VL - 28
SP - 165
EP - 171
JO - Stem Cell Research
JF - Stem Cell Research
SN - 1873-5061
ER -