Putative epidermal stem cell convert into corneal epithelium-like cell under corneal tissue in vitro
Putative epidermal stem cell convert into corneal epithelium-like cell under corneal tissue in vitro
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DOI：
10.1007/s11427-007-0006-4
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发表时间：
2007-02-01
期刊：
影响因子：
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通讯作者：
Cui GuangHui
中科院分区：
文献类型：
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作者：
Gao Nan;Wang ZhiChong;Cui GuangHui
恒河猴假定的表皮干细胞正在研究其在再生角膜上皮样细胞中的潜在用途，这可能为构建生物工程角膜提供实用的自体种子细胞来源。本研究的目的是探讨表皮干细胞向角膜上皮样细胞转分化的潜力。采用IV型胶原贴壁法分离恒河猴表皮干细胞。采用流式细胞术、免疫组织化学和RT-PCR方法检测细胞表面特异性标志物β 1、α 6整合素、K15、K1/K10、K3/K12和CD 71的表达。分离的细胞与人角膜缘基质和角膜上皮细胞共培养。共培养后，评价相同特异性标志物的表达，以鉴定由共培养条件引起的表达差异。在第2、4、6、8和10天分析共培养细胞中的K3/K12表达。条件培养基中的假定表皮干细胞用作对照。通过IV型胶原附着分离，推定的表皮干细胞在快速附着细胞中占主导地位。在共培养之前，恒河猴假定的表皮干细胞表达K15、α 6和β 1整合素，但不表达CD 71、K1/K10和K3/K12。共培养后，这些细胞表达K3/K12（角膜上皮细胞的标志物）、K15和β 1整合素，但不表达K1/K10。未共培养的细胞转化为表达K1/K10的终末分化细胞。这些结果表明，恒河猴假定的表皮干细胞可以转分化为角膜上皮样细胞，因此，可能有潜在的治疗应用作为自体种子细胞的生物工程角膜的建设。
Rhesus putative epidermal stem cells are being investigated for their potential use in regenerative corneal epithelium-like cells, which may provide a practical source of autologous seed cells for the construction of bioengineered corneas. The goal of this study was to investigate the potential of epidermal stem cells for trans-differentiation into corneal epithelium-like cells. Rhesus putative epidermal stem cells were isolated by type IV collagen attachment method. Flow cytometry analysis, immunohistology and RT-PCR were conducted to identify the expression of specific markers (beta 1, alpha 6 integrin, K15, K1/K10, K3/K12 and CD71) on the isolated rapid attaching cells. The isolated cells were cocultured with human corneal limbal stroma and corneal epithelial cells. After coculture, the expression of the same specific markers was evaluated in order to identify expression difference caused by the coculture conditions. K3/K12 expression was analyzed in coculture cells on day 2, 4, 6, 8 and 10. Putative epidermal stem cells in conditioned culture media were used as control. Putative epidermal stem cells were predominant in rapid attaching cells by type IV collagen attachment isolation. Before being cocultured, the rhesus putative epidermal stem cells expressed K15, alpha 6 and beta 1 integrin, but no CD71, K1/K10 and K3/K12. After coculture, these cells expressed K3/K12 (a marker of corneal epithelial cells), K15 and beta 1 integrin, but no K1/K10. Cells being not coculture converted into terminally differentiated cells expressing K1/K10. These results indicate that rhesus putative epidermal stem cells can trans-differentiate into corneal epithelium-like cells and, therefore, may have potential therapeutic application as autologous seed cells for the construction of bioengineered corneas.