脐带间充质干细胞增殖分化实验研究

目的:培养人脐带间充质干细胞并分析细胞的增殖及分化能力,为临床应用提供依据。方法:剥除新鲜脐带血管,取华通式胶,采用组织贴壁法培养细胞,将不同代数的细胞(P3~P6)采取两种方法(细胞计数法及MTT法)研究其增殖能力,并选取培养良好的P3代细胞进行分化实验研究。结果:人脐带间充质干细胞呈梭型流水状生长,并可在体外稳定扩增,细胞增殖呈‘S’型曲线,所培养细胞可以分化为成脂细胞、成骨细胞以及成软骨细胞。结论:人脐带可以分离培养间充质干细胞,间充质干细胞可在体外稳定扩增培养,细胞增殖呈现典型的‘S’型曲线,可将细胞增殖的对数生长期应用于临床,以便可以达到较好的效果,通过诱导分化表明所培养的细胞可以定向分化为成脂细胞、成骨细胞和成软骨细胞,表明所培养的细胞具有多向分化为不同细胞的潜能,其具有间充质干细胞的特性。

Objective:Human umbilical cord mesenchymal stem cells were cultured and studied differentiation and proliferation ability,providing a basis for clinical application.Methods:Blood vessels were stripped from fresh umbilical cord and cells taken from Wharton’s jelly were cultured by tissue adherent method.Two methods(cell counting method and MTT method)were adopted to study the proliferation ability from different generations(P3~P6)and the differentiation ability was studied from P3.Results:In vitro,Human umbilical cord mesenchymal stem cells presented with long spindle-shaped adherent growth and cell expansion was in steady state.Proliferation curve of cultured MSCs was“S”shape.Cultured cells could differentiate into adipocytes,osteoblasts and chondroblasts.Conclusion:Mesenchymal stem cells isolated from human umbilical cord could be amplified steadily in vitro.Proliferation curve of cultured MSCs was“S”shape,The growth phase of cell proliferation can be applied in clinic,In order to achieve good results.Umbilical Cord Mesenchymal Stem Cells could differentiate into adipocytes,osteoblasts and chondroblasts,and proved that Umbilical Cord Mesenchymal Stem Cells had the potential to differentiate into different cells and Characteristics of mesenchymal stem cells.