TY - JOUR
T1 - Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells
AU - Saari, Jaakko
AU - Siddique, Fatima
AU - Korpela, Sanna
AU - Mäntylä, Elina
AU - Ihalainen, Teemu O.
AU - Kaukinen, Katri
AU - Aalto-Setälä, Katriina
AU - Lindfors, Katri
AU - Juuti-Uusitalo, Kati
N1 - Funding Information:
Funding: This research was funded by Mary and George C. Ehrnrooth foundation (K.J.-U.), the Academy of Finland (S.K. decision number 323509, E.M. and T.O.I. decision number 332615, K.K. decision number 339183, K.L. decision number 314880), The Academy of Finland, Center of Excellence Body-on-Chip (K.A.-S.), Sigrid Juselius Foundation (K.K., K.A.-S., and K.L.), and the Competitive State Research Financing of the Expert Responsibility Area of Tampere University Hospital (K.K.).
Funding Information:
This research was funded by Mary and George C. Ehrnrooth foundation (K.J.-U.), the Academy of Finland (S.K. decision number 323509, E.M. and T.O.I. decision number 332615, K.K. decision number 339183, K.L. decision number 314880), The Academy of Finland, Center of Excellence Body-on-Chip (K.A.-S.), Sigrid Juselius Foundation (K.K., K.A.-S., and K.L.), and the Competitive State Research Financing of the Expert Responsibility Area of Tampere University Hospital (K.K.).
Publisher Copyright:
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/1
Y1 - 2022/1
N2 - The small intestinal epithelium has an important role in nutrition, but also in drug absorption and metabolism. There are a few two-dimensional (2D) patient-derived induced pluripotent stem cell (iPSC)-based intestinal models enabling easy evaluation of transcellular transport. It is known that animal-derived components induce variation in the experimental outcomes. Therefore, we aimed to refine the differentiation protocol by using animal-free components. More specifically, we compared maturation of 2D-cultured iPCSs toward small intestinal epithelial cells when cultured either with or without serum, and either on Geltrex or on animal-free, recombinant laminin-based substrata. Differentiation status was characterized by qPCR, immunofluorescence imaging, and functionality assays. Our data suggest that differentiation toward definitive endoderm is more efficient without serum. Both collagen-and recombinant laminin-based coating supported differentiation of definitive endoderm, posterior definitive endoderm, and small intestinal epithelial cells from iPS-cells equally well. Small intestinal epithelial cells differentiated on recombinant laminin exhibited slightly more enterocyte specific cellular functionality than cells differentiated on Geltrex. Our data suggest that functional small intestinal epithelial cells can be generated from iPSCs in serum-free method on xeno-free substrata. This method is easily converted to an entirely xeno-free method.
AB - The small intestinal epithelium has an important role in nutrition, but also in drug absorption and metabolism. There are a few two-dimensional (2D) patient-derived induced pluripotent stem cell (iPSC)-based intestinal models enabling easy evaluation of transcellular transport. It is known that animal-derived components induce variation in the experimental outcomes. Therefore, we aimed to refine the differentiation protocol by using animal-free components. More specifically, we compared maturation of 2D-cultured iPCSs toward small intestinal epithelial cells when cultured either with or without serum, and either on Geltrex or on animal-free, recombinant laminin-based substrata. Differentiation status was characterized by qPCR, immunofluorescence imaging, and functionality assays. Our data suggest that differentiation toward definitive endoderm is more efficient without serum. Both collagen-and recombinant laminin-based coating supported differentiation of definitive endoderm, posterior definitive endoderm, and small intestinal epithelial cells from iPS-cells equally well. Small intestinal epithelial cells differentiated on recombinant laminin exhibited slightly more enterocyte specific cellular functionality than cells differentiated on Geltrex. Our data suggest that functional small intestinal epithelial cells can be generated from iPSCs in serum-free method on xeno-free substrata. This method is easily converted to an entirely xeno-free method.
KW - 2D iPSC-SIEC
KW - Induced pluripotent stem cell
KW - IPSC
KW - IPSC-SIEC
KW - KO-SR
KW - Recombinant laminin
KW - Serum-free culture
KW - Small intestinal epithelial cell
U2 - 10.3390/ijms23031312
DO - 10.3390/ijms23031312
M3 - Article
AN - SCOPUS:85123310050
SN - 1661-6596
VL - 23
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 3
M1 - 1312
ER -