Toward Xeno-Free Differentiation of Human Induced Pluripotent Stem Cell-Derived Small Intestinal Epithelial Cells.
Jaakko SaariFatima SiddiqueSanna KorpelaElina MäntyläTeemu O IhalainenKatri KaukinenKatriina Aalto-SetäläKatri LindforsKati Juuti-UusitaloPublished in: International journal of molecular sciences (2022)
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.
Keyphrases
- endothelial cells
- stem cells
- induced apoptosis
- high glucose
- randomized controlled trial
- high resolution
- diabetic rats
- machine learning
- oxidative stress
- high throughput
- big data
- embryonic stem cells
- adipose tissue
- mesenchymal stem cells
- cell free
- skeletal muscle
- signaling pathway
- cell death
- insulin resistance
- cell therapy
- stress induced
- tissue engineering
- fluorescence imaging