小鼠胚胎干细胞与复合丝素膜的生物相容性研究

Biocompatibility Evaluation of Silk Fibroin with Mouse Embryonic Stem Cells in vitro

探索胚胎干细胞与丝素材料结合的可行性及生物相容性,建立一套在丝素膜上小鼠胚胎干细胞的体外培养体系。通过形态观察、MTT比色法、克隆形成率分析、碱性磷酸酶检测、免疫荧光染色、核型分析等方法,研究丝素膜对小鼠胚胎干细胞的黏附、生长情况、未分化状态的维持、胚胎干细胞特性、遗传、分化等方面的影响。结果显示,两者之间具有良好的生物相容性,丝素膜材料支持小鼠胚胎干细胞的体外无限扩增和未分化特征,并保持其正常核型。证实丝素材料可作为胚胎干细胞结合的良好生物材料。

The purpose of this study is to evaluate the biocompatibility of silk fibroin materials with mouse embryonic stem cells （ESCs） and to establish a culture system based on silk fibroin films. The growth of undifferentiated mouse ESC on silk fibroin films were investigated by MTT assay and colony efficiency analysis. The specific marker expression, inheritance and differentiation of mouse ESC on silk fibroin films were assessed by alkaline phosphatase （AKP） detection, immunocytochemistry, karyotyping and in vitro differentiation. The results showed that silk fibroin had good biocompatibility with mouse ESC and the established culture system supported the adhesion, growth and proliferation of ESC with pluripotency and normal karyotype. The silk fibroin materials support the growth and differentiation of embryonic stem ceils, thus providing important experimental information for tissue engineering.