Medical School of Soochow University, Suzhou, PR China.
Ophthalmic Res. 2012;48(2):82-88. doi: 10.1159/000331006.
Mesenchymal stem cells (MSCs) can play an active role in there construction of the ocular surface; however, the mechanisms by which the implanted MSCs exert their effects remain elusive. In the present study, we investigated the influences of MSCs on cell proliferation and growth factor expression of corneal limbal epithelial cells (LECs) in vitro. Rat LECs were either cocultured with rat MSCs or cultured in rat MSC condition edmedium. The harvested LECs were subjected to light microscopy, MTT assay, bromodeoxyuridine (BrdU)staining, flow cytometry and real-time quantitative RT-PCR analysis, respectively. Light microscopy showed that coculturing of LECs with MSCs significantly increased the proliferation of LECs. The MTT assay showed that MSC-conditioned medium significantly increased the cell viability of LECs compared to those incubated with plain medium. BrdU immunostaining and flow-cytometric analysis of the cell cycle showed that the cell proliferation rate of LECs cocultured with MSCs was significantly higher than that of LECs cultured alone. Real-time quantitative RT-PCR analysis showed that expression of epidermal growth factor in LECs cocultured with MSCs was significantly higher than that in LECs cultured alone. However, expression of basic fibroblast growth factor showed no difference under the two culture conditions. These results indicate that MSCs promote the survival and proliferation of LECs, and these effects may be mediated ina paracrine manner.
间充质干细胞（MSCs）在眼表的构建中可发挥积极作用；然而，植入的 MSCs 发挥作用的机制仍难以捉摸。在本研究中，我们研究了 MSCs 对体外角膜缘上皮细胞（LECs）的细胞增殖和生长因子表达的影响。将大鼠 LECs 与大鼠 MSCs 共培养或在大鼠 MSC 条件培养基中培养。分别对收获的 LECs 进行光镜检查、MTT 测定、溴脱氧尿苷（BrdU）染色、流式细胞术和实时定量 RT-PCR 分析。光镜检查显示，LECs 与 MSCs 共培养可显著增加 LECs 的增殖。MTT 测定显示，MSC 条件培养基可显著提高 LECs 的细胞活力，与普通培养基孵育的 LECs 相比。BrdU 免疫染色和细胞周期流式细胞术分析显示，与单独培养的 LECs 相比，与 MSCs 共培养的 LECs 的细胞增殖率显著更高。实时定量 RT-PCR 分析显示，与单独培养的 LECs 相比，与 MSCs 共培养的 LECs 中表皮生长因子的表达显著更高。然而，在两种培养条件下，碱性成纤维细胞生长因子的表达没有差异。这些结果表明，MSCs 促进 LECs 的存活和增殖，这些作用可能是通过旁分泌方式介导的。
