Saichanma Sarawut, Bunyaratvej Ahnond, Sila-Asna Monnipha
Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok 14140, Thailand.
Int J Ophthalmol. 2012;5(2):158-63. doi: 10.3980/j.issn.2222-3959.2012.02.08. Epub 2012 Apr 18.
The damage of human corneal cells encounter with the problem of availability of corneal cells for replacement. Limitation of the source of corneal cells has been realized. An attempt of development of corneal epithelial-like cells from the human skin-derived precursor (hSKPs) has been made in this study. Combination of three essential growth factors: epidermal growth factor (EGF), keratinocyte growth factor (KGF) and hepatocyte growth factor (HGF) could demonstrate successfully induction of hSKPs to differentiation into corneal cells.The induced cells expressed the appearance of markers of corneal epithelial cells as shown by the presence of keratin 3 (K3) by antibody label and Western blot assay. The K3 gene expression of induced hSKPs cells as shown by reverse transcription-polymerase chain reaction (RT-PCR) technology was also demonstrated. The presence of these markers at both gene and protein levels could lead to our conclusion that the directional transdifferentiation of hSKPs cells into corneal epithelial cells was successfully done under this cell induction protocol. The finding shows a newly available stem cell source can be obtained from easily available skin. Cells from autologous human skin might be used for corneal disorder treatment in future clinical application.
人类角膜细胞的损伤面临着用于替换的角膜细胞供应问题。人们已经认识到角膜细胞来源的局限性。本研究尝试从人皮肤来源的前体细胞(hSKPs)培养角膜上皮样细胞。三种重要生长因子的组合:表皮生长因子(EGF)、角质形成细胞生长因子(KGF)和肝细胞生长因子(HGF)能够成功诱导hSKPs分化为角膜细胞。通过抗体标记和蛋白质印迹分析,诱导细胞呈现出角膜上皮细胞标志物的表现,如角蛋白3(K3)的存在。逆转录聚合酶链反应(RT-PCR)技术也证实了诱导的hSKPs细胞中K3基因的表达。这些标志物在基因和蛋白质水平上的存在使我们得出结论,在这种细胞诱导方案下,hSKPs细胞成功地定向转分化为角膜上皮细胞。这一发现表明,可以从容易获取的皮肤中获得一种新的可用干细胞来源。来自自体人类皮肤的细胞可能在未来的临床应用中用于治疗角膜疾病。