Characterization of oral keratinocyte stem cells and prospects of its differentiation to oral epithelial equivalents.
Characterization of oral keratinocyte stem cells and prospects of its differentiation to oral epithelial equivalents.
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发表时间：
2010
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通讯作者：
B. Calenic;N. Ishkitiev;Ken Yaegaki;Toshio Imai;M. Costache;Mihaela Tovaru;Ș. Tovaru;I. Parlatescu
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作者：
B. Calenic;N. Ishkitiev;Ken Yaegaki;Toshio Imai;M. Costache;Mihaela Tovaru;Ș. Tovaru;I. Parlatescu
虽然口腔角质形成干细胞在组织稳态、伤口愈合和肿瘤形成中起着关键作用，但它们仍然难以鉴定和表征。本研究的具体目的是表征口腔角质形成细胞干细胞群分离使用磁性技术。材料与方法使用增殖相关标记物CD 71和α6β4整合素磁性分离从角化口腔粘膜获得的口腔人角化细胞。通过免疫荧光检查不同干细胞标志物：CD 44 H、Nestin、Nanog、Oct 3÷4、CD 117的表达。还测定了α6β4pos CD71阴性组分形成口腔上皮等同物的能力。结果经磁分离后获得3种不同的口腔角质形成细胞亚群：α6β4pos CD 71 neg、α6β4pos CD 71 pos和α6β4neg。我们的α6β4pos CD71neg干细胞部分对Oct 3 β 4、CD44 H和细胞角蛋白19呈阳性，而Nanog、Nestin和CD117表达缺失。同时，其他两种细胞组分α6β4pos CD71pos和α6β4neg对所有干细胞标志物均呈阴性。此外，α6β4pos CD71阴性组分能够再生分层良好且组织化良好的口腔上皮等同物。细胞角蛋白19和外皮蛋白在牙龈上皮中的分布反映了牙龈上皮的生物学特性。结论人牙龈α6β4pos CD71neg组分对一组干细胞标志物呈强阳性，并可形成口腔上皮等同物。也有人建议，磁系统可能是一个重要的工具，在获取口腔角质形成干细胞的研究。
OBJECTIVE Although oral keratinocyte stem cells play a key role in tissue homeostasis, wound healing, and neoplasia, they remain difficult to identify and characterize. The specific aim of the present study is to characterize an oral keratinocyte stem-cell population separated using a magnetic technique. MATERIAL AND METHODS Oral human keratinocytes obtained from keratinized oral mucosa were magnetically separated using a proliferation-related marker, CD71 and α6β4 integrin. The expression of different stem cell markers: CD44H, Nestin, Nanog, Oct 3÷4, CD117 was checked by immunofluorescence. The ability of α6β4pos CD71neg fraction to form oral epithelial equivalents was also assayed. RESULTS Three different oral keratinocyte subpopulations were obtained following magnetic separation: α6β4pos CD71neg, α6β4pos CD71pos and α6β4neg. Our α6β4pos CD71neg stem cell fraction was positive for Oct 3÷4, CD44H and cytokeratin 19 while Nanog, Nestin and CD117 expression was absent. At the same time, the other two cell fractions α6β4pos CD71pos and α6β4neg were negative for all stem cell markers. Also, α6β4pos CD71neg fraction was able to regenerate a well stratified and organized oral epithelial equivalent. The distribution of cytokeratin 19 and involucrin in the oral epithelial equivalent reflected the in vivo situation in oral gingival epithelium. CONCLUSIONS The human gingival α6β4pos CD71neg fraction was strongly positive for a panel of stem cell markers and could form oral epithelial equivalent. It is also suggested that a magnetic system may be an important tool in acquiring oral keratinocyte stem cells for research.