In vitro isolation and culture of rabbit bone marrow mesenchymal stem cells using directly adherent method
DING Chen1,ZHANG Zhongwen2,NIE Qinghu2
1.Graduate School Xuzhou Medical College, Xuzhou 221004, China; 2. 4th Orthopedics Department, General Hospital of Chinese People’s Armed Police Forces, Beijing 100039, China
Abstract：Objective To observe and identify the biological characteristics of rabbit bone marrow mesenchymal stem cells, and establish a set of simple , viable culture methods. Methods Rabbit bone marrow mesenchymal stem cells were extracted through adherent method directly, and use the inverted microscope to observe period morphology. The value-added level of the cells was assayed by MTT and the growth curve was drawn.The third passage of bone marrow mesenchymal stem cell phenotypes were measured by flow cytometry. Results Under the microscope,cells were small colony growth with shape of polygons when cells were cultured at the 5th day.Small colony confluent colony at 7-8 th day.The cells could be passaged at about 10th day.The second passage of cells showed long fusiform and fibroblast-like growth,that means marrow mesenchymal stem.The cell growth curve was S-shaped.Flow cytometry Results showed that the cells expressed CD 73,CD 90,CD 105, and did not express CD 34,CD 45. Conclusions The directly adherent method can get apundant, purified, and stable mesenchymal stem cells .
2014, Vol. 25 
