豚鼠骨髓间充质干细胞培养及成骨细胞定向分化

目的　分离和培养豚鼠骨髓间充质干细胞 ,研究其成骨细胞分化特性。方法　采用成年豚鼠用贴壁法分离骨髓间充质干细胞并传代培养。取第三代培养细胞做细胞生长曲线测定。用 β 甘油磷酸钠、地塞米松和维生素C诱导使其向成骨细胞分化。Gomori钙钴法作碱性磷酸酶染色。结果　全培养骨髓细胞 ,连续 3d换液后能得到均一的骨髓间充质干细胞。豚鼠骨髓间充质干细胞群体倍增时间为 6 1h。用 β 甘油磷酸钠、地塞米松和维生素C诱导可以使其成骨细胞分化。碱性磷酸酶染色阳性。结论　贴壁法可用于分离骨髓间充质干细胞。豚鼠骨髓间充质干细胞具有药物诱导定向成骨细胞分化的特性

Culture and differentiation of marrow mesenchymal stem cells in guinea pigs

Objective To separate and culture marrow mesenchymal stem cells in guinea pig bone marrow, and study the characteristics of guinea pig mesenchymal stem cells. Methods The mesenchymal stem cells were separated by adhering to the wall of plastic culture plate because of the different characteristics between mesenchymal stem cells and hemopoitic stem cells. We use the third passage cells to assay the growing curve of mesenchymal stem cells. β-glycerophosphate disodium salt hydrate, dexamethasome and vitamin C were used to make the guinea pig mesenchymal stem cells differentiate into osteoblasts. Results The marrow cells were totally cultured and the culturing solution was changed in 3 consecutive days. By the method we could obtain the mesenchymal stem cells. Population doubling time of guinea pig mesenchymal stem cells was about 61 h. The drugs could induce the differentiation of guinea pig stem cells into osteblasts. ALPase was expressed positively by the Gomori stain. Conclusion The methods of adhering to plate can separate the guinea stem cells. Guinea mesenchymal stem cells can be differentiated into osteoblasts by drug induction.