小鼠毛囊干细胞向心肌细胞的诱导分化及其鉴定

Induction and differentiation of mouse hair follicle stem cells into cardiomyocytes

目的探究小鼠毛囊隆突部分离培养的毛囊干细胞,诱导分化成为心肌细胞的能力。方法显微镜下剥离结构完整的小鼠毛囊,只保留隆突部,Ⅳ型胶原黏附法分离、培养隆突部细胞,对其进行形态学和分子免疫荧光检测;第二代细胞的单细胞克隆通过有限稀释法纯化,并筛选K15、K19强阳性的细胞;加入心肌细胞诱导液进行诱导分化并做心肌细胞标志物肌红蛋白的免疫荧光鉴定。结果小鼠毛囊隆突部干细胞经成心肌诱导后,细胞呈现典型的心肌细胞样形态,K15、K19呈阳性;诱导后的细胞能特异表达心肌细胞的标志物肌红蛋白。结论小鼠毛囊干细胞有分化成为心肌细胞的能力。

Objective To investigate the ability of hair follicle stem cells isolated and cultured from mouse hair follicle bulge to induce and differentiate into cardiomyocytes.Methods The dissected hair follicles with complete structure were peeled off under the microscope,and only the bulge was retained.Cells of the bulge were isolated and cultured by the typeⅣcollagen adhesion method,and their morphology were observed and molecular immunofluorescence were done.The single cell clone of the second generation cells was pontificated by limiting dilution method,K15 and K19 strong positive cells were screened.Cardiomyocyte induction solution was added into the culture medium to induce and differentiation,and immunofluorescence identification was done to indentify mygotobin which was the marker of cardiomyocytes.Results The stem cells of the hair follicle of the mice were induced into cardiomyocytes,and the cells showed typical cardiomyocyte-like morphology with positive expression of K15 and K19.Myoglobin,the marker of cardiomyocyte,were specifically expressed after induction.Conclusion Mouse hair follicle stem cells have the ability to differentiate into cardiomyocytes.