凝集性生长和非凝集性生长人毛乳头细胞线粒体的结构及膜电位比较

目的比较凝集性生长和非凝集性生长人毛囊毛乳头细胞(DPC)的线粒体数量、分布、结构和功能特点。方法采用二步酶消化法分离人毛囊毛乳头后,体外培养DPC。利用还原型线粒体特异性荧光探针MitoTrackerRedCM-H2Xros,结合激光扫描共聚焦显微镜和透射电镜观察两种不同生长状态DPC线粒体的分布、数量及结构特征;利用四氯四乙基苯并咪唑基羰花青碘化物(JC-1)检测线粒体膜电位。结果体外培养DPC生长良好;与非凝集性生长DPC相比,凝集性生长DPC线粒体数量较多,整个胞浆内可见,且集中分布在粗面内质网附近,线粒体外形较不规则,嵴较发达,线粒体膜电位明显较高。结论凝集性生长DPC和非凝集性生长DPC的线粒体数量、分布、结构和功能不同,说明两者在能量消耗、细胞代谢及蛋白质合成功能方面存在差异。

Comparative study on structure and membrane potentials of mitochondria in cultured human dermal papilla cells under aggregative and non-aggregative conditions

Objective To observe the number,distribution,structure and function of mitochondria in cultured human dermal papilla cells (DPCs) under aggregative and non-aggregative conditions.Methods DPCs were separated from human hair follicles by two-step enzymatic digestion and cultured in DMEM medium.Mitochondrion-selective stain,laser scanning confocal microscopy and transmission electron microscopy were used to observe the number,distribution and structure of mitochondria.Mitochondria membrane potential (Δψ_m)was detected by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocy-anine iodide (JC-1).Results DPCs grew well in the culture.Compared with the DPCs with non-aggregative behavior,those with aggregative behavior had more mitochondria,which were visible throughout the cytoplasm and concentrated near the rough endoplasmic reticulum,with irregular shape,well-developed cristae and higher membrane potentials.Conclusions The number,distribution,structure and function of mitochondria are different between the DPCs under aggregative condition and those under non-aggregative condition,which suggests there are differences in energy consumption,cell metabolism and protein synthesis between the two conditions of DPCs.