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旨在探索鸡胚胎干细胞(ES细胞)体外定向诱导的多向分化潜能。分离鸡X期胚盘细胞,体外培养传代,经鉴定后,采用地塞米松(DEX)、β-甘油磷酸钠(β-GP)和维生素C(VitC)诱导ES细胞向成骨细胞分化;采用维甲酸(RA)、3-异丁基-1-甲基黄嘌呤(IBMX)诱导ES细胞向神经元样细胞分化;采用DEX、胰岛素、IBMX诱导ES细胞向脂肪细胞分化,比较不同浓度组合诱导剂的诱导效果,分别用Von Kossa’s染色法、甲苯胺蓝染色法、免疫组化法、油红O染色法和RT-PCR鉴定诱导产生的细胞。结果显示,ES细胞被诱导3～21d后,分化为成骨细胞,诱导率为40%～72%;被诱导4～7d后,分化为神经元样细胞,诱导率为71%～84%;被诱导2～21d后,分化为脂肪细胞,油红O染色阳性,并表达PPARγ基因。结果表明,在体外条件下,ES细胞可被特异的化学物质定向诱导分化为成骨细胞、神经元样细胞和脂肪细胞,具有多向分化潜能。
The purpose of this study was to explore the multi-directional differentiation potential of chicken embryonic stem cells (ES cells) induced in vitro. Isolation of chicken x stage blastodermal cells, in vitro culture passage, after identification, dexamethasone (DEX), β-glycerophosphate (β-GP) and vitamin C (VitC) induced differentiation of ES cells into osteoblasts; Retinoic acid (RA) and 3-isobutyl-1-methylxanthine (IBMX) induced differentiation of ES cells into neuron-like cells. Differentiation of ES cells into adipocytes was induced by DEX, insulin and IBMX. The induced cells were identified by Von Kossa’s staining, toluidine blue staining, immunohistochemistry, oil red O staining and RT-PCR, respectively. The results showed that ES cells differentiated into osteoblasts after induction for 3 ~ 21 days, the induction rate was 40% -72%; after being induced for 4 ~ 7 days, they differentiated into neuron-like cells with the induction rate of 71% -84%; After being induced for 2 ~ 21 days, they differentiated into adipocytes, stained with Oil Red O, and expressed PPARγ gene. The results show that under in vitro conditions, ES cells can be differentially differentiated into osteoblasts, neuron-like cells and adipocytes by specific chemical substances and have the potential of multi-directional differentiation.