The Effect Of Various Concentrations Of Allogenic Platelet-rich Plasma Exacting Solution On Adipose-derived Stem Cells In Vitro

Objective：To explore the effect of various concentrations of allogenic platelet-rich plasma（PRP） extracting solution on adipose-derived stem cells（ADSCs） in vitro. In order toprovide an experimental evidence for wound healing by allogenic PRP extractingsolution.Methods：The adipose tissue derived from SD rats were isolated, cultured and passaged;Cells harvested above were cultured respectively in adipose-, osteo-and neuro-lineages intrducing culture solution. Oil red O stain for adipose-induction, Von Kossastain for osteo-induction and specific immunofluorescence stain for neuro-inductionwere performed to identify the differentiation of these cells. The rats’ blood wereextracted and prepared by three times’ centrifugation for making PRP and differentconcentrations of PRP （6.67%,3.35%, and1.67%volume fractions） were prepared byDMEM/F12. The third passage of ADSCs were divided into4groups: the ADSCswere cultured with6.67%volume fractions of PRP in group A, with3.35%volumefractions of PRP in goup B, and with1.67%volume fractions of PRP in goup C, andwith10%fetal bovine serum in complete medium in group D. The effects of the fourgroups on the proliferative activities of the ADSCs were detected by MTT after24and48hours to reflect cell proliferation activity. The data was analyzed by SPSS17.0.Results：The harvested cells appeared to be adherent and fibroblast-like cells, which weresuccessfully obtained and culured from rat’s adipose tissue. The cells could differe-ntiate into adipocyte, osteocyte and neurocyte by adipose-induced, osteo-induced,neuro-induced. It is proved that the cells have the potential of differentiating intoadipose-,osteo-and neuro-lineage cells and that the harvested cells are ADSCs. Thenumber of the platelet in PRP was621.05×10⁹/L, which was5times as many as thatin the whole blood (309.26×10¹⁰/L). The OD value assayed by MTT after24hours in Group A、B、C、D were spectively0.4343±0.0843,0.3516±0.0766,0.2581±0.0600,0.2592±0.0736and that after48hours were1.0186±0.4436,0.7417±0.1097,0.3678±0.0342,0.3957±0.1062. The OD value in Group A and B were significantlyhigher than that in goup C and D, while the OD value in Group A were higher thanthat in group B, showing significant differences （P<0.05）. However, there was nosignificant difference between group C and D （P>0.05）.Conclusion：A large number of ADSCs could be abtained by collagenase digestion andprimary culture. In vitro, the proliferation of SD rat’s ADSCs could be enhanced byallogenetic PRP extracting solution, which is expected to accelerate wound repairing.