Isolation of oral epithelial progenitors using collagen IV
Isolation of oral epithelial progenitors using collagen IV
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DOI：
10.1111/j.1601-0825.2007.01390.x
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发表时间：
2008-07-01
期刊：
影响因子：
3.8
通讯作者：
Yamane, G-y
中科院分区：
文献类型：
--
作者：
Igarashi, T.;Shimmura, S.;Yamane, G-y
目的：虽然口腔粘膜上皮干细胞被认为存在于基底层，但这种细胞尚未被分离出来。我们分离了兔口腔上皮祖细胞群，其中含有假定的干细胞。材料和方法：将兔颊黏膜上皮细胞粘附在涂有IV型胶原的培养皿上10分钟至16小时。使用BrdU、Ki-67、整合素β 1、整合素α 6和角蛋白13（集落形成效率（CFE））评估单个细胞群体的特性。结果：黏附于iv型胶原包被培养皿10 min内的细胞BrdU掺入量增加约6倍，Ki-67、整合素α 6、整合素β 1均有强表达。有趣的是，角蛋白13表达微弱。口腔上皮细胞中快速贴壁细胞的CFE与其他细胞群比较显著。结论：兔口腔上皮细胞可通过黏附IV型胶原进行分离，根据祖细胞特性分离效果更好。具有干细胞特性的推定祖细胞在10分钟内最有效地收获。我们的分离方法将成为分离上皮干细胞用于再生医学的有用工具。
OBJECTIVE: Although oral mucosal epithelial stem cells are thought to reside in the basal layer, such cells have not yet been isolated. We isolated a population of rabbit oral epithelial progenitor cells containing putative stem cells.MATERIALS AND METHODS: Epithelial cells harvested from rabbit buccal mucosa were allowed to adhere to dishes coated with collagen IV for periods ranging from 10 min to 16 h. The properties of individual cell populations were evaluated using BrdU, Ki-67, integrin beta 1, integrin alpha 6 and keratin 13 using colony forming efficiency (CFE).RESULTS: Cells that adhered to collagen IV-coated dishes within 10 min were enriched about sixfold in terms of BrdU incorporation, Ki-67, integrin alpha 6 and integrin beta 1 were strongly expressed. Interestingly, keratin 13 was faintly expressed. The CFE of rapidly adherent cells among oral epithelial cells was significant compared with other cell populations.CONCLUSIONS: These results suggested that rabbit oral epithelial cells could be isolated by depending on adhesiveness to collagen IV, especially when segregated according to progenitor cell properties. Putative progenitor cells with stem cell properties were most effectively harvested within 10 min. Our separation procedure should be a useful tool with which to isolate epithelial stem cells for regenerative medicine.