论文部分内容阅读
本研究体外培养并用抗角质蛋白K3/K12鼠单抗(AE5)鉴定人角膜缘干细胞(HLSCs),采用脂质体转染法将pEGFP-bFGF基因转染培养的HLSCs,48 h后荧光显微镜下可见特异性的绿色荧光,基因转染率为20%～30%,同时制作NaOH细胞损伤模型,研究转染后的细胞对损伤的抵抗作用,转染后损伤的细胞存活状况高于未转染损伤组(P<0.05),而细胞凋亡及坏死率明显低于未转染损伤组(P<0.05)。pEGFP-bFGF基因能够转染培养的HLSCs,其表达产物对碱损伤的HLSCs具有保护作用,初步探讨了基因工程联合组织工程技术治疗眼表疾病的可行性。
In this study, human limbal stem cells (HLSCs) were identified by using anti-keratin K3 / K12 mouse monoclonal antibody (AE5) in vitro. HUVECs were transfected into HLSCs by lipofection method. After 48 h, Specific green fluorescence, gene transfection rate of 20% to 30%, at the same time making NaOH cell injury model to study the transfection of cells after injury resistance, cell survival after transfection damage was higher than non-transfected Group (P <0.05), while the apoptosis and necrosis rate were significantly lower than the untransfected group (P <0.05). The pEGFP-bFGF gene can be transfected into cultured HLSCs, and its expression product has a protective effect on alkali-injured HLSCs. The feasibility of gene engineering combined with tissue engineering to treat ocular surface diseases has been discussed.