O 13. Effect of different concentrations of fetal calf serum on the in vitro culture of rat bone marrow mesenchymal stem cells
Objective: To establish a method of isolating and culturing rat bone marow mesenchymal stem cells (MSCs) in vitro and to analyse their phenotypical properties.
Methods: Rat bone marrow cells were separated and expanded by adherence culture. The primary MSCs were respectively cultured by adhering in L-DMEM containing 15% and 10% FBS. After passage, all cells were cultured by adhering in L-DMEM containing 10% FBS. The morphologic features, the growth curve, and the surface markers including CD44, CD45 and CD90 of MSCs were observed.
Results: MSCs of primary culture in 15% FBS had more rapid proliferation, earlier colony confluence, and shorter time for passage primary (about 8 days) and passage 1 (about 5 days) than in 10% FBS. The growth curves of passage 3 MSCs showed a similar total cell number between the two groups (P>0.05). MSCs after passage in both groups expressed CD44 and CD90 but not CD 45.
Conclusions: The primary MSCs cultured in L-DMEM containing 15% FBS can provide MSCs in shorter period compared with the 10% FBS by adherence culture.
Key words
Mesenchymal stem cells; fetal bovine serum; mice; cell culture