Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells

Jaakko Saari; Fatima Siddique; Sanna Korpela; Elina Mäntylä; Teemu O. Ihalainen; Katri Kaukinen; Katriina Aalto-Setälä; Katri Lindfors; Kati Juuti-Uusitalo

doi:10.3390/ijms23031312

Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells

Jaakko Saari
, Fatima Siddique
, Sanna Korpela
, Elina Mäntylä
, Teemu O. Ihalainen
, Katri Kaukinen
, Katriina Aalto-Setälä
, Katri Lindfors
, Kati Juuti-Uusitalo^*

^*Corresponding author for this work

TAYS Heart Centre

Research output: Contribution to journal › Article › Scientific › peer-review

4 Citations (Scopus)

45 Downloads (Pure)

Abstract

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.

Original language	English
Article number	1312
Journal	International Journal of Molecular Sciences
Volume	23
Issue number	3
DOIs	https://doi.org/10.3390/ijms23031312
Publication status	Published - Jan 2022
Publication type	A1 Journal article-refereed

Funding

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.). 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.).

Keywords

2D iPSC-SIEC
Induced pluripotent stem cell
IPSC
IPSC-SIEC
KO-SR
Recombinant laminin
Serum-free culture
Small intestinal epithelial cell

Publication forum classification

Publication forum level 1

ASJC Scopus subject areas

Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry

Access to Document

10.3390/ijms23031312Licence: CC BY

ijms-23-01312-v2Final published version, 4.75 MBLicence: CC BY

https://urn.fi/URN:NBN:fi:tuni-202205104628Licence: CC BY

iPS Cells
Aalto-Setälä, K. (Manager)
Facility/equipment: Research infrastructure

Cite this

@article{dd45c26695d64e57aad02055c643df07,

title = "Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells",

abstract = "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.",

keywords = "2D iPSC-SIEC, Induced pluripotent stem cell, IPSC, IPSC-SIEC, KO-SR, Recombinant laminin, Serum-free culture, Small intestinal epithelial cell",

author = "Jaakko Saari and Fatima Siddique and Sanna Korpela and Elina M{\"a}ntyl{\"a} and Ihalainen, \{Teemu O.\} and Katri Kaukinen and Katriina Aalto-Set{\"a}l{\"a} and Katri Lindfors and Kati Juuti-Uusitalo",

note = "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: {\textcopyright} 2022 by the authors. Licensee MDPI, Basel, Switzerland.",

year = "2022",

month = jan,

doi = "10.3390/ijms23031312",

language = "English",

volume = "23",

journal = "International Journal of Molecular Sciences",

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publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",

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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 -

Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells

Abstract

Funding

Keywords

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