两种方法提取的人血清对体外培养人真皮成纤维细胞促增殖作用的比较研究

Effect of human serum extracted by two different methods on human dermal fibroblast growth in vitro

目的使用两种不同方法提取人血清,研究对比得出最快速、廉价、安全而有效的富含生长因子的人血清提取方法,以促进其在临床上的应用。方法分离并培养人真皮成纤维细胞。分别使用全血凝固后离心法和Anitua法提取人血清。测定各血清样本中PDGF-AB及TGF-β1的含量。分别使用含有10%胎牛血清(FBS)和10%人血清(HS)的DMEM培养基培养成纤维细胞,比较不同培养基对细胞形态、活力、增殖的影响。结果各组细胞形态正常,增殖活力良好。B组(全血凝固后离心组)细胞的数量及活力均明显高于其他各组(P<0.05),且该法获得的血清中生长因子含量最高(P<0.05),与生长曲线及MTT曲线所示结果一致。结论全血凝固后离心法提取的HS内所含生长因子浓度最高,对细胞生长促进作用最大,提取方法简单、廉价、有效。

Objective To explore the optimal method for extracting human serum that retains rich growth factors.Methods Human serum was isolated by centrifugation of coagulated whole blood or by Anitua＇s method,and the concentrations of PDGF-AB and transforming growth factor-β1（TGF-β1） in the serum samples were measured.Human dermal fibroblasts were cultured in the presence of 10% feral bovine serum or 10% human serum obtained,and the cell morphology,viability and proliferative activity of the cells were evaluated.Results The fibroblasts grew well in all the media with good viability.The cells cultured in the presence of human serum isolated by centrifugation of coagulated whole blood,which had the richest content of growth factors,showed the greatest cell number and cell viability among the groups（P0.05）,a result consistent with the growth curve and MTT curve.Conclusion Centrifugation of coagulated whole blood retains high contents of growth factors in human serum to better promote cell growth,and is simple,cost-effective and most efficient for serum isolation.