大鼠骨髓间充质干细胞体外可诱导分化为角膜上皮细胞

目的:探讨骨髓间充质干细胞 (mesenchymal stem cell,MSC) 分化为角膜上皮细胞的可塑性及其重建角膜上皮的可能性.方法:用密度梯度离心法结合贴壁培养法分离纯化大鼠骨髓MSC,经体外与角膜基质细胞共培养诱导分化,免疫荧光法检测角膜上皮细胞特异标志物K12的表达.结果:体外培养的大鼠骨髓MSC表现出很强的增殖潜能,原代培养的骨髓MSC CD29免疫荧光染色阳性,CD34和CD45为阴性,符合骨髓MSC的特征.MSC与角膜基质细胞共培养1wk后大部分细胞分化为间质细胞,少部分细胞形态上相对偏小,免疫荧光检测这部分细胞表达角膜上皮细胞特异性标志角蛋白K12.结论:体外培养的MSC在角膜基质细胞的诱导下可横向分化为角膜上皮细胞.

Study on transdifferentiation of rat's mesenchymal stem cells into corneal epithelial cells in vitro

AIM:To explore the plasticity of transdifferentiation of mesenchymal stem cells (MSC) into corneal epithelial cells. METHODS:MSC of adult rats that were isolated and purified by density gradient centrifugation combined with an attachment culture method. The transdifferentiation of MSC into corneal epithelial cells were induced in vitro by co-cultured with corneal stromal fibroblasts. The expression of K12 on MSCs which is expressed specially in corneal epithelial cells was identified by immunofluorescent staining. RESULTS:MSC cultured in vitro showed great potential of proliferation. CD29 was positive,and CD34,CD45 were negative in cultured cells,indicating that the cultured cells were MSC. After co-cultured with corneal stromal fibroblasts for one week,MSC expressed K12,indicating that they had been transdifferentiated into corneal epithelial cells. CONCLUSION:The data suggests that MSC have the potential to transdifferentiate into corneal epithelial cells in vitro.