中文摘要：

【目的】通过类胚体介导体外自由分化与诱导分化,验证所建立的牛诱导性多能干细胞(Bovine induced pluripotent stem cells,biPSCs)能否分化为外胚层的神经细胞。【方法】将biPSCs悬浮培养以制备类胚体,通过类胚体介导使biPSCs在添加全反式维甲酸(RA)和β巯基乙醇(β-Me)诱导体系中分化为神经细胞,比较其诱导效率;提取不同诱导体系的分化细胞与类胚体的总RNA,利用RT-PCR方法鉴定分化细胞中神经细胞特异性基因的表达。【结果】培养的biPSCs集落呈球形,中央隆起,周围界限清晰,与胚胎干细胞集落形态类似。biPSCs碱性磷酸酶(AP)染色为阳性,并表达SSEA-4干细胞特异性表面蛋白。biPSCs通过类胚体介导分化,在未添加化学诱导物的条件下,可自由分化为Nestin、GFAP与NSE阳性神经细胞,其阳性细胞率分别为(15.14±1.13)%,(6.25±0.35)%和(5.45±0.62)%;biPSCs在RA诱导组中分化的神经细胞的数量最多,诱导后表达Nestin、GFAP和NSE细胞的阳性率分别为(49.56±2.33)%,(16.58±1.28)%和(13....

英文摘要：

【Objective】Bovine induced pluripotent stem cells(biPSCs)have been generated in the previous study.This study was conducted to verify the differentiation potential of neural lineage cells from the resulting biPSCs under different induction conditions.【Method】The resulting biPSCs were cultured using the hanging drop and suspension culture method for embryoid body(EBs)formation,and EBs were transferred into 0.1% gelatin-coated 24-well plates for attached culture.The attached cells were divided into three induc...