| Project/Area Number |
23792364
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Surgical dentistry
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| Research Institution | Niigata University (2012)
Kagoshima University (2011) |
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Principal Investigator |
SAITO Yoko 新潟大学, 医歯学総合病院, 助教 (30404487)
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| Project Period (FY) |
2011 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2011: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Keywords | 小児歯科学 / 再生医学 / バイオリサイクル / 歯 / iPS細胞 / iＰＳ細胞 |
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| Research Abstract |
Feeder cells are generally required for establishing and maintaining embryonic stem (ES) cells/inducible pluripotent stem (iPS) cells. Mouse embryonic fibroblasts (MEFs) isolated from fetuses and STO mouse stromal cell line are the most widely used cells for this purpose. The aim of this study was to determine which cells are suitable for establishing iPS cells from human deciduous tooth dental pulp cells (HDDPCs) and supernumerary teeth (SNT). Primarily cultured HDDPCs and SNT (5 × 104) wereco-transfected with 3 plasmids containing human OCT3/4, SOX2/KLF4, or LMYC/LIN28 together with pmaxGFP (used as a reporter) by using the NeonR Transfection system, a novel electroporation method involving a capillary and wire-type electrode. They were then cultured in a medium specified for maintaining ES cells for 15 days. Colonies were collected by trypsinization and re-seeded onto mitomycin C-treated MEFs or STO cells. The colonies were serially passaged for up to 26 passages. During this period, colony morphology was assessed to determine whether they exhibit ES-like morphology and alkaline phosphatase activity to evaluate the state of HDDPC reprogramming. HDDPCs maintained on MEFs were transformed to iPS cells with several undifferentiated properties, but those on STO cells failed. Established HDDPC-derived iPS cells grown on MEFs were successfully maintained on STO cells without loss of their pluripotent nature. Our results indicate that HDDPCs can be reprogrammed using reprogramming factor, and that MEFs are better than STO cells as feeder cells for establishing and maintaining HDDPC-derived iPS cells, suggesting that the choice of feeders is a key factors enabling efficient generation of iPS cells.
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