Generation of kidney tubular organoids from human pluripotent stem cells

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

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number38353
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 2016 Dec 16

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Organoids
Pluripotent Stem Cells
Kidney
Proteins
Cultured Cells
Kidney Tubules
Glycogen Synthase Kinase 3
Stem Cell Research
Bioengineering
Antibodies
Nephrons
Cell Transplantation

ASJC Scopus subject areas

  • General

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Generation of kidney tubular organoids from human pluripotent stem cells. / Yamaguchi, Shintaro; Morizane, Ryuji; Honma, Koichiro; Monkawa, Toshiaki; Suzuki, Sayuri; Fujii, Shizuka; Koda, Muneaki; Hiratsuka, Ken; Yamashita, Maho; Yoshida, Tadashi; Wakino, Shu; Hayashi, Koichi; Sasaki, Junichi; Hori, Shingo; Itoh, Hiroshi.

In: Scientific Reports, Vol. 6, 38353, 16.12.2016.

Research output: Contribution to journalArticle

Yamaguchi, S, Morizane, R, Honma, K, Monkawa, T, Suzuki, S, Fujii, S, Koda, M, Hiratsuka, K, Yamashita, M, Yoshida, T, Wakino, S, Hayashi, K, Sasaki, J, Hori, S & Itoh, H 2016, 'Generation of kidney tubular organoids from human pluripotent stem cells', Scientific Reports, vol. 6, 38353. https://doi.org/10.1038/srep38353
Yamaguchi, Shintaro ; Morizane, Ryuji ; Honma, Koichiro ; Monkawa, Toshiaki ; Suzuki, Sayuri ; Fujii, Shizuka ; Koda, Muneaki ; Hiratsuka, Ken ; Yamashita, Maho ; Yoshida, Tadashi ; Wakino, Shu ; Hayashi, Koichi ; Sasaki, Junichi ; Hori, Shingo ; Itoh, Hiroshi. / Generation of kidney tubular organoids from human pluripotent stem cells. In: Scientific Reports. 2016 ; Vol. 6.
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