Analysis of side population cells as keratinocyte stem cells.
| Project/Area Number |
16591102
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Dermatology
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| Research Institution | Ehime University |
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Principal Investigator |
KAMEDA Kenji Ehime University, Integrated Center for Sciences, lecturer, 総合科学研究支援センター, 講師 (60363264)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRAKATA Yuji Ehime University, Graduate School of Medicine, lecturer, 大学院医学系研究科, 講師 (50226320)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | stem cells / SP-cells / cell sorter / keratinocyte |
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| Research Abstract |
Cultured skin is widely used for skin defects such as chronic ulcer, burns and so on. To develop gene therapy using cultured skin, it is necessary to introduce foreign genes into keratinocyte stem cells. Keratinocyte stem cells have been isolated using cell surface markers, however this method requires multiple steps for staining, and cell damage might occur because of the use of antibodies. Side population (SP) cells was firstly reported as stem cell isolation method for hematopoietic cells. The purpose of this study is to investigate whether SP cells exist in keratinocytes and SP cells are keratinocyte stem cells or not. Normal human skin was obtained from plastic surgery. Epidermis was separated from dermis by dispase treatment, single cells of keratinocytes were obtained by trypsin treatement. Keratinocytes were collected and cultured under serum free condition. Cells were stained with Hoechst 33342, then Hoechst Red and Blue expression was analyzed by cell sorter. SP cells exist in normal keratinocytes, and the population rate is 1-5% in total cells. Next we performed microarray analysis. SP cells as well as main population (MP) cells were sorted and total RNA was extracted. Using gene chip carrying 27000 genes, about 10 genes were significantly up-or down-regulated. Finally we investigated proliferative potency of SP cells. SP cells and MP cells were sorted and cultured separately. When cells reached subconfluent, cells were passaged. SP cells had higher proliferative potency by 3-4 passages, however MP cells grew well after 5 passages. These data indicate that SP cells might be the transit amplifying cells rather than stem cells, however further studies are needed.
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Report
(4 results)
Research Products
(15 results)
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[Journal Article] Hematopoietic stem cells prevent hair cell death after transient cochlear ischemia through paracrine effects2007
Author(s)
Yoshida T, Hakuba N, Morizane I, Fujita K, Cao F, Zhu P, Uchida N, Kameda K, Sakanaka M, Gyo K, HataR
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Journal Title
Neuroscience 145
Pages: 923-930
Description
「研究成果報告書概要(和文)」より
Related Report
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[Journal Article] Hematopoietic stem cells prevent hair cell death after transient cochlear ischemia through paracrine effects2007
Author(s)
Yoshida T, Hakuba N, Morizane I, Fujita K, Cao F, Zhu P, Uchida N, Kameda K, Sakanaka M, Gyo K, HataR.
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Journal Title
Neuroscience 145
Pages: 923-930
Related Report
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[Journal Article] TAK1 is essential for differentiation and the prevention of apoptosi in epidermis2006
Author(s)
Sayama K, Hanakawa Y, Nagai H, Shirakata Y, Dai X, Hirakawa S, Tokumaru S, Tohyama M, Yang. L, Sato S, Shizuo A, Hashimoto K
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Journal Title
J Biol Chem 281
Pages: 22013-22020
Description
「研究成果報告書概要(和文)」より
Related Report
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[Journal Article] TAK1 is essential for differentiation and the prevention of apoptosis in epidermis.2006
Author(s)
Sayama K, Hanakawa Y, Nagai H, Shirakata Y, Dai X, Hirakawa S, Tokumaru S, Tohyama M, Yang L, Sato S, Shizu A, Hashimoto K.
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Journal Title
J Biol Chem. 281
Pages: 22013-20
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] TAK1 is essential for differentiation and the prevention of apoptosis in epidermis.2006
Author(s)
Sayama K, Hanakawa Y, Nagai H, Shirakata Y, Dai X, Hirakawa S, Tokumaru S, Tohyama M, Yang L, Sato S, Shizuo A, Hashimoto K
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Journal Title
J Biol Chem 281
Pages: 22013-20
Related Report
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