小鼠胚胎干细胞结合丝素材料向神经细胞分化的实验研究

以小鼠胚胎干细胞（ES）为种子细胞，使用改良的4-/4+ RA方案，诱导小鼠ES细胞在丝素材料上向神经细胞分化，探讨丝素材料对其生长、黏附、分化等情况的影响。将小鼠ES细胞悬浮培养4 d得到的拟胚体（EBs）分别接种到经丝素膜和明胶包被的培养皿上进行诱导，比较不同材料上EBs的贴壁率及向神经元分化的比率。结果表明EBs在明胶和柞蚕丝素蛋白膜（TSF）上贴壁较快，平均贴壁率为90.3%和84.4%，在桑蚕丝素蛋白膜（SF）上贴壁较慢，贴壁率低，仅为38.5%，同时三者神经元的分化比率均能达到40%以上，无明显差异。通过以上实验，我们得出，TSF有望成为小鼠ES细胞向神经细胞分化的支架材料。

Differentiation of mouse embryonic stem cells into neural cells on silk fibroin scaffolds

The feasibility of embryonic stem cells to combine with silk fibroin for neural differentiation was invaluable and unknown. We suspended the ES cells into EBs and then transferred them to three different substrates-coated 35 mm dishes including gelatin, Bombyx mori silk fibroin (SF) and Tussah silk fibroin (TSF) to identify the adherence and proportion of ES cells-derived neurons under these three substrates. The results showed that silk fibroin as a reliable biomaterial which can support EBs’ adherence and differentiated into euron-like cells compared with control group, all the proportion of β-Ⅲ-Tubulin positive cells is approximately 40%. The results further proved that the adhesion ability of cells on SF is poorer than on the control group and TSF group, which also showed that TSF is an excellent biomaterial for the neural differentiation of ES cells. This study may provide important experimental information for tissue engineering, in which ES cells-derived neuron cells and silk fibroin materials are scaffolds, and also offer a source for cell therapy research of neurodegenerative disease.