兔骨髓基质细胞向成骨细胞分化过程的超微形态结构观察  

Morphological and ultrastructural observation of bone marrow stromal cells in the process of osteogenic differentiation in rabbits

作　　者：杨成林[1] 毕郑钢[1] 刘勇 王玉学[1] 祁权[1] 

机构地区：[1]哈尔滨医科大学附属一院骨科,哈尔滨市150001 [2]哈尔滨市第五医院骨科

出　　处：《中华显微外科杂志》2007年第4期281-284,I0011,共5页Chinese Journal of Microsurgery

基　　金：黑龙江省科委十五攻关项目(GBO1C1234-01);黑龙江省教育厅资助课题(11511215);黑龙江省卫生厅资助课题(2005-190)

摘　　要：目的观察并探讨骨髓基质细胞向成骨细胞诱导分化过程中细胞超微形态结构的变化及相互联系。方法抽取成年兔髂骨骨髓,分离、培养骨髓基质细胞,1周后分别采用含诱导因子的条件培养液和小含诱导因子的标准培养液继续培养,2周后收集细胞,应用透射电镜观察其超微结构;同时分别将上述两组细胞与煅烧骨体外复合并在原培养液中继续培养,分别于复合1、7、14d取出,用扫描电镜观察细胞表面超微形态。结果透射电镜下见诱导前的骨髓基质细胞浆少,核大,细胞器不发达,处于早期幼稚阶段,而诱导2周后的细胞核小、胞浆多,细胞器丰富,处于较为成熟阶段。扫描电镜下见复合培养2周内诱导前后的骨髓基质细胞均由圆盘形向多角形、条带状相互融合、重叠生长,诱导2周后的骨髓基质细胞间可见大量胶原纤维等基质成分以及钙盐颗粒,而诱导前的细胞始终未见基质分泌及钙盐颗粒。结论成骨诱导后,骨髓基质细胞内细胞器等超微结构极为丰富,细胞表面及细胞间有大量细胞外基质分泌和钙盐沉积。透射电镜和扫描电镜可动态观察骨髓基质细胞向成骨细胞分化过程中细胞超微形态结构的变化。Objective To observe the morphological and ultrastructural changes of bone marrow stromal cells（BMSCs） in the process of osteogenic differentiation, and analyze their relationship. Methods BMSCs were isolateded from rabbit bone marrow, grown in culture. After 1 week, BMSCs were cultured in standard medium without osteogenic agents and in experimental culture medium with osteogenic agents respectively. Two weeks later, BMSCs were collected, and the ultrastructure of which were observed using transmission electronic microscope （TEM）. Then induced and noninduced BMSCs were seeded into ceramic bovine bone（CBB） to form cell-scaffold constructs, and cell-scaffold constructs cultured continually in the original medium respectively were examined with scaning electronic microscope（SEM） after 1,7,14 days. Results The larger nucleus, lesser cytoplasm, rare organelles were seen inside noninduced cells, which were immature. However the lesser nucleus, greater cytoplasm, a great deal of organelles were easily seen inside induced cells, which were mature. And then BMSCs changed from round dish shape to star polygan or tape form, which fused and lapped over each other in 2 weeks under SEM. On day 14th, a lot of collagen filaments and calcium granules were found on the surface of induced cells or in the intercellular space, no cell matrix and calcium deposition were observed among the noninduced cells. Conclusion A great of organelles, cell matrix such as collgen filaments and calcium granules were observed after BMSCs were induced osteogein differentiation. TEM and SEM could continually observe the morphological and ultrastuctural changes of BMSCs in the process of osteogenic differentiation.

关 键 词：骨髓基质细胞 成骨细胞 超微结构 形态学 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]