Co-Culture of Mesenchymal-Like Stromal Cells Derived From Human Foreskin Permits Long Term Propagation and Differentiation of Human Embryonic Stem Cells
Co-Culture of Mesenchymal-Like Stromal Cells Derived From Human Foreskin Permits Long Term Propagation and Differentiation of Human Embryonic Stem Cells
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DOI：
10.1002/jcb.23052
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发表时间：
2011-05-01
影响因子：
4
通讯作者：
Bhonde, Ramesh
中科院分区：
文献类型：
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作者：
Mamidi, Murali Krishna;Pal, Rajarshi;Bhonde, Ramesh
在决定人胚胎干细胞（human embryonic stem cells，hESC）成功分化和扩增的不同参数中，饲养层起着最重要的作用。以人间充质基质细胞（hMSCs）和人包皮成纤维细胞（HFFs）形式存在的人饲养层为根除动物源性hESC培养体系奠定了基础。在这项研究中，我们探讨了人包皮来源的间充质样基质细胞（HF-MSCs）的潜力，以支持自我更新和hESC的多能性。发现从人包皮分离的MSC对标准浓度和持续时间的丝裂霉素-C治疗具有抗性。生长模式、基因分析（Oct-4、Nanog、Sox-2、雷克斯-1）、细胞骨架蛋白表达（波形蛋白、巢蛋白）和向脂肪细胞、软骨细胞和骨细胞的三系分化潜能证实了它们的间充质基质细胞状态。此外，HF-MSC对CD 105、CD 166、CD 73、CD 44、CD 90、SSEA-4呈阳性，而对CD 34、CD 45、HLA-DR细胞表面标记呈阴性，并被发现表现出BM-MSC样特征。与HF-MSC饲养细胞共培养的hESC系显示出预期的多能转录因子Oct-4、Nanog、Sox-2、GDF-3、雷克斯-1、STELLAR、ABCG 2、Dppa 5、hTERT的表达;表面标志物SSEA-4、TRA-1-81，并在长期传代期间保持其细胞遗传学稳定性。这些新型饲养细胞还改善了hESC胚状体（EB）的形成，产生了代表三个种系的细胞类型。这种培养系统有可能帮助开发用于再生医学和药物筛选的临床级hESC。此外，我们设想包皮可以作为一个有价值的来源替代骨髓间充质干细胞的具体治疗应用。J.细胞。112：1353-1363，2011. (C)2011 Wiley-Liss，Inc.
Among the different parameters governing the successful derivation and expansion of human embryonic stem cells (hESC), feeder layers play the most important role. Human feeders in form of human mesenchymal stromal cells (hMSCs) and human foreskin fibroblasts (HFFs) lay the foundation for eradication of animal-derived hESC culture system. In this study we explored the potential of human foreskin derived mesenchymal like stromal cells (HF-MSCs) to support self renewal and pluripotency of hESC. The MSCs isolated from human foreskin were found to be resistant to standard concentrations and duration of mitomycin-C treatment. Growth pattern, gene profiling (Oct-4, Nanog, Sox-2, Rex-1), cytoskeletal protein expression (vimentin, nestin) and tri-lineage differentiation potential into adipocytes, chondrocytes and osteocytes confirmed their mesenchymal stromal cell status. Further, the HF-MSCs were positive for CD105, CD166, CD73, CD44, CD90, SSEA-4, and negative for CD34, CD45, HLA-DR cell-surface markers and were found to exhibit BM-MSC-like characteristics. hESC lines cocultured with HF-MSC feeders showed expression of expected pluripotent transcription factors Oct-4, Nanog, Sox-2, GDF-3, Rex-1, STELLAR, ABCG2, Dppa5, hTERT; surface markers SSEA-4, TRA-1-81 and maintained their cytogenetic stability during long term passaging. These novel feeders also improved the formation of embryoid bodies (EBs) from hESC which produced cell types representing three germ layers. This culture system has the potential to aid the development of clinical-grade hESCs for regenerative medicine and drug screening. Further, we envisage foreskin can serve as a valuable source of alternative MSCs for specific therapeutic applications. J. Cell. Biochem. 112: 1353-1363, 2011. (C) 2011 Wiley-Liss, Inc.