孤雌胚胎干细胞向成纤维细胞和表皮细胞的定向诱导及构建双层组织工程皮肤的研究

项目名称： 孤雌胚胎干细胞向成纤维细胞和表皮细胞的定向诱导及构建双层组织工程皮肤的研究

项目编号： No.31300797

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 崔继红

作者单位： 西北大学

项目金额： 23万元

中文摘要： 种子细胞是皮肤组织工程研究和应用的关键。构建双层TES需要用到表皮细胞和成纤维细胞,但原代培养的表皮细胞和成纤维细胞来源有限，而且细胞质量不稳定。胚胎干细胞具有很强的增殖能力和多向分化潜能，其来源的表皮细胞已被成功应用于皮肤缺损修复，但应用会受到伦理学和免疫排斥反应的限制。本研究将孤雌胚胎干细胞诱导分化为表皮细胞和成纤维细胞，进行双层TES构建研究。首先扩增小鼠pESCs并向表皮细胞和成纤维细胞诱导分化，对其表型特征进行系统观察，重点观察细胞的角质形成能力和FGF、EGF、PDGF、VEGF、TGF、IGF等生长因子的表达水平；进而用pESCs来源的表皮细胞和成纤维细胞构建双层TES，观察双层TES的生物学特征；最后进行异体移植，观察皮肤缺损修复、表型维持、排斥反应等指标，深入评价这种新型双层TES的应用前景。

中文关键词： 孤雌胚胎干细胞；成纤维细胞；种子细胞；组织工程皮肤；

英文摘要： Skin regeneration is an important area of research in the tissue engineering (TE) field, especially for massive skin loss cases, where current treatments are yet not capable of inducing permanent satisfying skin regeneration. To achieve an effective healing, skin TE products must attach well to the wound bed, be supported by new vasculature, integrate with the surrounding host tissues, be nonimmunogenic, and be capable of self regeneration with minimum scar tissue, with reduced patient pain and discomfort, and yet importantly, manufactured with a good cost-benefit ratio. Cultured skin began when methods for harvesting keratinocytes from patients and proliferating the cells in vitro became available. While engineered skin substitutes represent significant advances in wound care, their use is not routine because of their high cost, limited effectiveness, and their inability to reconstitute skin appendages.Embryonic stem cells(ESCs) are harvested from the inner cell mass of the blastocyst and have complete potential to differentiate into all the primary germ layers such as ectoderm, mesoderm, and endoderm. ESCs could offer a virtually unlimited source of cells for use in tissue repair and regeneration. Stepwise approach to generate non-tumorigenic and lineage-restricted chondrogenic cells from ESCs under defined

英文关键词： pESCs；fibroblasts；seed cells；TES；