血小板裂解液对人间充质干细胞生物学特性的影响

Effect of platelet lysate on the biological characteristics of human mesenchymal stem cells

背景:血小板裂解液是自体或异体血小板富集物经裂解后获得的产物,不仅去除了残余的细胞结构,降低了免疫原性,而且保留了其中的多种生长因子,有研究表明它可替代传统培养基中的胎牛血清,对间充质干细胞进行体外扩增。目的:观察血小板裂解液对人骨髓间充质干细胞和脂肪间充质干细胞生物学特性的影响,为临床细胞治疗及再生医学提供科学的实验数据。方法:采集新鲜血液,经冻融获取血小板裂解液;采集健康成人骨髓和脂肪组织,分别采用密度梯度离心法和Ⅰ型胶原酶消化法获得骨髓间充质干细胞和脂肪间充质干细胞;对血小板裂解液培养后的骨髓间充质干细胞和脂肪间充质干细胞的细胞形态、细胞表型、分化能力、增殖能力、集落形成能力和细胞因子分泌水平进行检测。结果与结论:应用血小板裂解液在体外成功地培养扩增了骨髓间充质干细胞和脂肪间充质干细胞,两种细胞的细胞形态、细胞表型、集落形成能力、细胞因子分泌水平、成软骨、成骨和成脂肪分化能力等生物学特性无明显差异。脂肪间充质干细胞的累积群倍数高于骨髓间充质干细胞(P<0.05),表明脂肪间充质干细胞具有更强的增殖能力,在体外血小板裂解液培养体系下更适宜进行大规模扩增。

BACKGROUND：Platelet lysate has been known as a kind of lysate of autologous or allogeneic platelet-rich products. It not only removes the residual cell structure,reduces immunogenicity,but also retains many growth factors. Platelet lysate has been suggested as a substitute for fetal bovine serum to expand mesenchymal stem cells in vitro.OBJECTIVE： To observe the effects of platelet lysate on biological characteristics of human bone marrow mesenchymal stem cells and adipose mesenchymal stem cells,and provide some experimental data for clinical cell therapy and regenerative medicine.METHODS： Platelet lysate was prepared by repeated freezing and thawing from fresh blood. Healthy adult bone marrow and adipose tissue were collected. Human bone marrow mesenchymal stem cells and adipose mesenchymal stem cells were obtained by density gradient centrifugation and type I collagenase digestion. We tested the morphology,cell phenotype,differentiation characteristics,proliferation capacity,colony forming ability and the level of cytokine secretion of bone marrow mesenchymal stem cells and adipose mesenchymal stem cells after cultured with platelet lysis.RESULTS AND CONCLUSION： Bone marrow mesenchymal stem cells and adipose mesenchymal stem cells were successfully cultured in vitro using platelet lysate. There were no significant differences in morphology,cell phenotype,colony forming ability and the level of cytokine secretion,and chondrogenic,osteogenic and adipogenic capacities between bone marrow mesenchymal stem cells and adipose mesenchymal stem cells. Adipose mesenchymal stem cells had a high cumulative population doublings than bone marrow mesenchymal stem cells（P〈0.05）. These findings suggest adipose mesenchymal stem cells had a stronger proliferative ability,and are more suitable for large-scale expansion in vitro cultivation system of platelet lysate compared with bone marrow mesenchymal stem cells.