The Use of Human Mesenchymal Stem Cell-Derived Feeder Cells for the Cultivation of Transplantable Epithelial Sheets
The Use of Human Mesenchymal Stem Cell-Derived Feeder Cells for the Cultivation of Transplantable Epithelial Sheets
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DOI：
10.1167/iovs.08-2262
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发表时间：
2009-05-01
影响因子：
4.4
通讯作者：
Tsubota, Kazuo
中科院分区：
文献类型：
--
作者：
Omoto, Masahiro;Miyashita, Hideyuki;Tsubota, Kazuo
目的。目的：探讨人骨髓间充质干细胞作为饲养细胞来源培养可移植的角膜上皮细胞的效果。方法：人骨髓间充质干细胞(MASCs)在含10%血清的改良Eagle‘s培养液中培养，用逆转录聚合酶链式反应(RT-PCR)检测MASCs中细胞因子的表达。将人角膜缘上皮细胞与MASCs或3T3饲养层细胞共培养，比较集落形成效率(CFE)。将角膜缘上皮细胞培养在气液界面的MASCs或3T3饲养层细胞上，进行细胞角蛋白3(K3)、K15、p63α和ABCG2免疫组织化学染色。将兔角膜缘上皮细胞片与MASC饲养层细胞复合培养，移植到角膜缘缺损兔的眼表。结果MASC饲养层细胞表达角质形成细胞生长因子、肝细胞生长因子和N-钙粘素。人角膜缘上皮细胞的CFE在MASC组和3T3饲养层组相似。用表达K3、K15、p63α和ABCG2的MASC饲养层细胞成功地培养了分层细胞片。结论：MASC来源的饲养层细胞适合于上皮层的构建，避免了潜在危险的异种饲养层细胞的使用。(投资眼科VS科学。DOI：10.1167/iovs.08-2262.
PURPOSE. To report the efficacy of human bone marrow-derived mesenchymal stem cells as a source of feeder cells for the cultivation of transplantable corneal epithelial cell sheets.METHODS. Human mesenchymal stem cells (marrow adherent stem cells; MASCs) were cultured in alpha-modified Eagle's medium with 10% serum and were treated with mitomycin C. Expression of cytokines in MASCs was confirmed by reverse transcription-polymerase chain reaction. Human limbal epithelial cells were cocultured with MASCs or 3T3 feeder cells to compare colony-forming efficiency (CFE). Limbal epithelial cells were cultured on MASCs or 3T3 feeder cells at the air-liquid interface to allow stratification, and stratified epithelial sheets were analyzed by immunohistochemistry against cytokeratin 3 (K3), K15, p63 alpha, and ABCG2. Rabbit limbal epithelial cell sheets were cultivated with MASC feeder cells and transplanted to the ocular surface of the limbal-deficient rabbits. Epithelial grafts were observed by slit lamp microscopy for 4 weeks and then evaluated by histology and immunohistochemistry against K3 and K4.RESULTS. MASC feeder cells expressed keratinocyte growth factor, hepatocyte growth factor, and N-cadherin. The CFE of human limbal epithelial cells was similar in MASC and 3T3 feeder groups. Stratified cell sheets were successfully cultivated with MASC feeder cells expressing K3, K15, p63 alpha, and ABCG2. Transplanted epithelial sheets regenerated the corneal phenotype in limbal-deficient rabbits.CONCLUSIONS. MASC-derived feeder cells are suitable for the engineering of epithelial sheets, avoiding the use of potentially hazardous xenologic feeder cells. (Invest Ophthalmol Vis Sci. 2009;50:2109-2115) DOI:10.1167/iovs.08-2262