人骨膜细胞体外培养的实验

In vitro culture of human periosteal cells

目的:观察体外培养的人骨膜细胞的形态学特征。方法:实验于2001-01/2003-10广东省粤北人民医院骨科和医学动物实验室完成。采用常规细胞培养法进行人骨膜细胞体外培养,并采用倒置显微镜和电子显微镜观察骨膜细胞生长情况、形态特征及超微结构。结果:①人骨膜培养活细胞光镜下形态:早期新生细胞呈梭形,饱满透明,立体感强;分裂期呈立方形或短柱状;后期由长梭形逐渐变成宽梭和不规则形,分泌功能旺盛。②人骨膜细胞电镜下超微结构:胞体呈梭形,膜面有少量微绒毛状突起,粗面内质网丰富,有大量异噬体等次级溶酶体,线粒体体积较小、基质致密有少量嵴。具有原代细胞生物形态学特征。结论:人骨膜成骨细胞可经体外培养大量增殖,且细胞传代进化后,其形态学结构与原代细胞无明显差异,可用于细胞移植。

AIM： To observe the morphological characteristics of human periosteal cells cultured in vitro. METHODS： The experiment was carried out in the Department of Orthopaedics and Laboratory of Medical Animal of Yuedei People＇s Hospital from January 2001 to October 2003. The human periosteal cells were cultured by the conventional method in vitro, and the growth, morphological characteristics and uhrastrueture of the periosteal cells were observed under inverted microscope optics and electronic microscope. RESULTS： ① Form of the survived human periosteal cells under light microscope： The neonatal periosteal cells showed shuttle, satiation, clarity and third dimensions at the early stage, and then became cube or puncheon at the split stage, and gradually became wide shuttle and irregular shape with a lot of secretion at telophase. ② Ultrastructure of the human periosteal cells under electronic microscope： The bodies of the ceils showed fusiform, and there were a bit of microvilli on membranous wall and abundant endoplasmic reticula. The volume of mitochondria whose matrix had a few cristallin was less than that of normal perioteal cells. The cells possessed the morphological characteristics as the primary ones. CONCLUSION： Human periosteal cell can be greatly cultivated and proliferated in vitro. Their cellular configuration has no distinct difference from the primary cells after anagenesis, the cultured cells can be used in transplantation.