A Refined Culture System for Human Induced Pluripotent Stem Cell-Derived Intestinal Epithelial Organoids

Publication date: Available online 7 December 2017 Source:Stem Cell Reports Author(s): Yu Takahashi, Shintaro Sato, Yosuke Kurashima, Tomohisa Yamamoto, Shiho Kurokawa, Yoshikazu Yuki, Naoki Takemura, Satoshi Uematsu, Chen-Yi Lai, Makoto Otsu, Hiroshi Matsuno, Hideki Osawa, Tsunekazu Mizushima, Junichi Nishimura, Mikio Hayashi, Takayuki Yamaguchi, Hiroshi Kiyono Gut epithelial organoids are routinely used to investigate intestinal biology; however, current culture methods are not amenable to genetic manipulation, and it is difficult to generate sufficient numbers for high-throughput studies. Here, we present an improved culture system of human induced pluripotent stem cell (iPSC)-derived intestinal organoids involving four methodological advances. (1) We adopted a lentiviral vector to readily establish and optimize conditioned medium for human intestinal organoid culture. (2) We obtained intestinal organoids from human iPSCs more efficiently by supplementing WNT3A and fibroblast growth factor 2 to induce differentiation into definitive endoderm. (3) Using 2D culture, followed by re-establishment of organoids, we achieved an efficient transduction of exogenous genes in organoids. (4) We investigated suspension organoid culture without scaffolds for easier harvesting and assays. These techniques enable us to develop, maintain, and expand intestinal organoids readily and quickly at low cost, facilitating high-throughput screening of pathogenic factors and cand...
Source: Stem Cell Reports - Category: Stem Cells Source Type: research