3次元構造を有する腎臓再生を実現するための基盤となる研究を行うため, ヒトES/iPS細胞から尿細管細胞への分化誘導方法の検討をkidney specific protein(KSP)を指標に行った。GSK3β阻害剤および尿細管分化培地を用いた2段階のシンプルな培養方法により尿細管細胞へ分化誘導し, 抗KSPモノクローナル抗体を用いて尿細管細胞の純化を行った。KSP陽性細胞は尿細管細胞の機能を有し, Wnt4およびマウス胎児腎との3次元培養により, 尿細管管腔構造を形成した。これらの結果は, KSP陽性細胞が純化された尿細管細胞であることを示し, 今後の腎臓再生のツールとして期待される。
Methods to purify specific types of cells from differentiated human pluripotent stem cells (hPSCs) would be useful to obtain pure populations of specific types of kidney cells. We report a simple two-step differentiation protocol to generate kidney tubular organoids from hPSCs with direct purification of KSP-positive cells using anti-KSP antibody. We differentiated hPSCs into mesoderm cells using a glycogen synthase kinase-3βinhibitor, subsequently cultured cells in renal epithelial growth medium to induce KSP+ cells. We purified KSP+ cells using flow cytometry with anti-KSP antibody, and cultured KSP+ cells in 3D Matrigel, which formed tubular organoids in vitro. 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. This approach is useful for generating cells of kidney tubular lineage cells that would be usable as a source for kidney regeneration.

研究種目 : 若手研究(B)
研究期間 : 2014～2015
課題番号 : 26860643
研究分野 : 腎臓再生