Effect of autologous platelet-rich plasma on proliferation of rabbit bone marrow mesenchymal stem cells

Abstract

Objective To establish a new method to culture bone marrow mesenchymal stem cells (BMSCs),which can avoid ethic controversy,transmitting diseases,causing immunological antigenicity problems and so on,induced by using heterogeneous serum,and meet the demand on various growth factors.Methods Platelet-rich plasma (PRP) was prepared by using Haemospasia from rabbit' s ear central artery.Punctured and sampled bone marrow from rabbit' s iliac bone,and collected BMSCs,which divided into PRP group,FBS group and serum-free control group,and cultured in 10％ autologous PRP,10％ FBS and serum-free respectively,combined with DMEM-F12 medium.To observe cells morphology,detect proliferation and draw growth curves,cells surface antigen and ALP activity of each group.Results BMSCs of 3 groups adhered to wall after inoculating about 24 hours,presented round and changed into fusiform,polygonal,and irregular forms gradually.The growth curves of 3 groups were “S” shape,PRP and FBS groups proliferated more rapidly than serum-free control group (P ＜ 0.0l ),and PRP group more than FBS group ( P ＜ 0.05 ).CD44 of the 4th passage BMSCs of PRP and FBS group was 88.73 ％ and 85.66％,and CD90 was 84.32％ and 87.58％,both expressed positively; but CD34 was 12.38％ and 14.76％,and CD45 was 10.47％ and 11.82％,both expressed negatively.The ALP activity among FBS group and serum-free control group from the 1st to 5th passage was similar,but the activity of PRP group increased as passaging,and which of 4th and 5th (6.42 ±0.89),(7.02 ±0.92) U/100 ml was significantly higher than 1st and 2nd (4.27 ±0.74),(4.70 ±0.76) U/100 ml (P ＜0.01 ),and higher than the 4th and 5th passage cells of PRP and FBS group(P ＜ 0.01 ).Conclusion Autologous PRP could promote BMSCs proliferating effectively,the cells morphology,proliferation activity and surface antigen cultured in PRP was similar as in FBS; and the ALP activity increased when it was passaged,which indicated the PRP had the potential to induce BMSCs to osteoblast. Key words: BMSCs; Platelet; Plasma; Tissue engineering