2012 Fiscal Year Final Research Report
Established efficiency of iPS cells derived from impacted supernumerary tooth considering bio-recycle
| 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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| Keywords | 小児歯科学 |
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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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[Presentation] Choice of feeders is important for the preparation of iPS cells from primarily cultured human deciduous tooth dental pulp cells2013
Author(s)
村上智哉,齊藤一誠,稲田絵美,岩瀬陽子,長谷川大子,窪田直子,松本祐子,大島邦子,岡暁子,山崎要一,早崎治明
Organizer
第51回日本小児歯科学会大会
Place of Presentation
岐阜市
Year and Date
20130523-24
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