The Effect Of Different Concentrations Of Fetal Bovine Serum On Bone Marrow Mesenchymal Stem Cell Purity And Cycle

Objective To compare the influence of three kinds of complete media with0.10,0.15,0.20fetal bovine serum(FBS) on purity and cycle of rat bone marrowmesenchymal stem cells (BMSCs)cultured in vitro and to seek suitable FBSconcentration for the cultivation of the stem cells.Methods SD rats were executed by cervical dislocation method and used whole bonemarrow adherence to isolate rat BMSCs. Experiment was divided into A,B,C,3groups.Compared the expression of CD45, CD29, CD90, CD44in the three groups in2,3,4,5passages(P2, P3, P4, P5); added osteogenesis induced liquid in group A of P3, observehe result of calcium nodule formation with alizarin red stain; the cells of P3in group Awere digested and cultured in three different concentration of complete culture mediafor four days, measuring cell cycle in24,48,72,96h by flow cytometry instrument.BMSCs of P3were collected and inoculated to6pieces of96-well plates, thenvaccinated with complete media with0.10,0.15,0.20FBS in every plate, one cultureplate was taken out for optical density(OD) meaturement every day with CCK-8.Results CD45was negative, CD29, CD90, CD44were positive. The difference ofBMSCs surface makers cultured in the three kinds of complete media was bigger in thefirst two passages, but the difference was less in P3, P4, all could obtain pure BMSCs inP4relatively; calcium nodule formation were found in groupA of P3after osteogenicinduced; according to the results of the cell cycle at the same time, G0/G1phase: with the increase of concentration of fetal bovine, G0/G1phase reduced and had nodiference in A, B, C groups; G2/M phase: there was difference between them after24h（P<0.05，F=12.412）, but with the extension of time, the differences disappeared; Sphase: there was no difference in the three groups; S+G2/M phase increased with theconcentration of FBS. According to the result of cell vitality, the OD of ABC threegroups increased in turn in24h, and there were difference（P<0.05，F=5.002）, but withthe extension of time, there was no obvious difference between them.Conclusions The three kinds of culture media in P4can obtain pure BMSCs, cellcycle and vitality show three complete medias could promote the growth of BMSCs,there was no difference between them. culture media with0.10FBS could satisfy theisolation and amplification of BMSCs, in order to obtain pure BMSCs in the short term,using culture media with0.15and0.10FBS in the primary culture and subculturerespectively.