Generation of kidney tubular organoids from human pluripotent stem cells

Shintaro Yamaguchi, Ryuji Morizane, Koichiro Homma, Toshiaki Monkawa, Sayuri Suzuki, Shizuka Fujii, Muneaki Koda, Ken Hiratsuka, Maho Yamashita, Tadashi Yoshida, Shu Wakino, Koichi Hayashi, Junichi Sasaki, Shingo Hori, Hiroshi Itoh

研究成果: Article査読

21 被引用数 (Scopus)

抄録

Recent advances in stem cell research have resulted in methods to generate kidney organoids from human pluripotent stem cells (hPSCs), which contain cells of multiple lineages including nephron epithelial cells. Methods to purify specific types of cells from differentiated hPSCs, however, have not been established well. For bioengineering, cell transplantation, and disease modeling, it would be useful to establish those methods to obtain pure populations of specific types of kidney cells. Here, we report a simple two-step differentiation protocol to generate kidney tubular organoids from hPSCs with direct purification of KSP (kidney specific protein)-positive cells using anti-KSP antibody. We first differentiated hPSCs into mesoderm cells using a glycogen synthase kinase-3β inhibitor for 3 days, then cultured cells in renal epithelial growth medium to induce KSP+ cells. We purified KSP+ cells using flow cytometry with anti-KSP antibody, which exhibited characteristics of all segments of kidney tubular cells and cultured KSP+ cells in 3D Matrigel, which formed tubular organoids in vitro. The formation of tubular organoids by KSP+ cells induced the acquisition of functional kidney tubules. KSP+ cells also allowed for the generation of chimeric kidney cultures in which human cells self-assembled into 3D tubular structures in combination with mouse embryonic kidney cells.

本文言語English
論文番号38353
ジャーナルScientific reports
6
DOI
出版ステータスPublished - 2016 12 16

ASJC Scopus subject areas

  • 一般

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