小鼠角膜上皮祖细胞系TKE2的表型研究 |
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引用本文: | 瞿杨洛娃,林辉,耿志鑫,何卉,刘祖国,李炜.小鼠角膜上皮祖细胞系TKE2的表型研究[J].中华细胞与干细胞杂志(电子版),2014(1):17-20. |
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作者姓名: | 瞿杨洛娃 林辉 耿志鑫 何卉 刘祖国 李炜 |
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作者单位: | 厦门大学眼科研究所福建省眼科与视觉科学重点实验室,厦门361102 |
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基金项目: | 基金项目:国家重大科学研究计划项目(2013CB967003);国家自然科学基金面上项目(81270977);国家卫生和计划生育委员会科研基金项目(WK3-FJ-26);福建省高等学校新世纪优秀人才支持计划项目 |
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摘 要: | 目的:观察小鼠角膜上皮祖细胞系TKE2在扩增以及分化状态下的角蛋白及干细胞标志物的表达情况。方法小鼠角膜上皮祖细胞系TKE2在无血清培养基Keratinocyte-SFM (KSFM)以及含10﹪胎牛血清(FBS)的DMEM培养基中培养,约70﹪融合时进行角蛋白10、12、14、15、16(K10、K12、K14、K15、K16)以及Connexin43、ABCG2的免疫荧光染色,以及Ki67、P63、PCNA的免疫细胞化学染色。结果无血清培养状态下的TKE2细胞呈克隆样生长,克隆内所有细胞呈ABCG2、K14、Ki67、PCNA以及P63阳性,K15阳性细胞散在分布,K16阳性细胞呈片状分布于克隆中央区,K10、K12以及Connexin43染色为阴性。在含有10﹪胎牛血清的DMEM中培养2 d后,细胞明显增大, ABCG2、K15、P63、Ki67以及PCNA转为阴性,克隆内只有少量细胞呈K16、K14阳性染色, K10、K12、Connexin43仍为阴性。结论 TKE2细胞具有角膜上皮干细胞特性,可以作为角膜缘上皮干细胞表型维持和分化诱导研究的良好工具。
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关 键 词: | 上皮 角膜 干细胞 细胞增殖 细胞分化 |
The phenotype study of murine corneal epithelial progenitor cell line TKE2 |
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Authors: | Qu Yangluowa Lin Hui Geng Zhixin He Hui Liu Zuguo Li Wei |
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Institution: | (Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Hsual Science, Xiamen 361102, China) |
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Abstract: | Objective To investigate the expression of stem cell markers and cytokeratins in murine corneal epithelial progenitor cell line TKE2 under proliferation and differentiation culture condition. Methods TKE2 cells were cultured in Keratinocyte-SFM (KSFM) and Dulbecco’s modified eagle medium (DMEM) with 10﹪fetal bovine serum (FBS). The cultures were terminated after TKE2 reached to 70﹪confluence. The expression patterns of stem cell markers (ABCG2, Ki67, P63, PCNA), proliferation related cytokeratins (K14, K15, K16), corneal terminal differentiation markers (K12, Connexin43) and epidermal epithelial specific marker (K10) on the TKE2 cells cultured under two conditions were evaluated by immunofluorescence and immunochemistry. Results In the KSFM medium, TKE2 cells could form uniform clones at the low seeding density. The expression of ABCG2, Ki67, PCNA, P63 and K14 were present in all cells of TKE2 clones. K15 were discretely expressed by TKE2 cells, while only sporadic cells in the central area of the clones expressed K16. After cultivation for 2 days in DMEM medium with 10﹪FBS, the cells became flat and enlarged. The expression of ABCG2, K14, K15, P63, Ki67 and PCNA were completely lost. K14 and K16 were weakly expressed in a small cluster of cells inside the TKE2 clones. K10, K12 and Connexin43 were negative in two culture conditions. Conclusions TKE2 processed the major characteristic of murine limbal stem cells. TKE2 cells may act as the potential cell model to study the mechanism of stemness maintenance as well as stem cell differentiation in limbal epithelium. |
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Keywords: | Epithelium Corneal Stem cell: Cell proliferation Cell differentiation |
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