诱导鸡胚精原干细胞向成骨细胞分化的研究

为了探讨体外培养的鸡胚精原干细胞(SSCs)的多向分化潜能,取孵化18~20 d的鸡胚睾丸,无菌获取SSCs,体外培养传代,通过地塞米松、β-甘油磷酸钠、维生素C诱导鸡胚SSCs向成骨细胞分化,钙结节Von Kossa s法、改良的钙钴法及免疫组化法进行成骨细胞鉴定。结果显示SSCs被诱导15~21 d后分化为成骨细胞,诱导形成率为75%~80%;诱导后的细胞Von Kossa s染色可见细胞间布满黑色颗粒,提示有矿化基质沉积;改良钙钴法碱性磷酸酶染色胞浆呈深棕色或深黑色,免疫组化法染色细胞呈阳性。因此可以得出在不同的诱导条件下,SSCs体外可被定向诱导分化为成骨细胞,证明其具有多向分化潜能。

Study on differentiation of chicken spermatogonial stem cells into osteoblasts in vitro

The current project is conducted to investigate the capability of spermatogonial stem cells(SSCs) differentiating into osteoblasts in vitro.SSCs were isolated from chicken embryo testis on 18th~20th hatching days,then cultured and subcultured in vitro.SSCs were induced into osteoblasts by desamethason,glycerol 2-phosphate disodium salt hydrate and vitamin C and examined by Von Kossa's stain,cytochemistry staining and immunohistochemistry staining.The results showed that osteoblasts were differentiated from SSCs after 15~21 days of induction and with 75%~80% of positive cells efficiency.Black granules were distributed in the differentiated cells by Von Kossa's staining,suggesting mineral cumulation in the cells.Our results were also confirmed by ALP and immunohistochemistry staining.Together,our results indicated that the chicken SSCs could be differentiated into osteoblasts,suggesting SSCs have the capability of multipotential differentiation in vitro.