| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Outline of Final Research Achievements |
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.
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