摘要
以青年男性股骨骨膜为材料，采用植块组织培养技术，在ＤＭＥＭ／Ｆ１２加１５％灭活小牛血清培养基中，建立了人成骨细胞体外培养模型，追踪观察了成骨细胞在体外培养５０天中的形态变化，并通过碱性磷酸酶（ＡＬＰ）染色技术，对培养２０天的原代细胞超微结构进行了酶细胞化学研究。结果显示：（１）成骨细胞为成纤维细胞型，细胞贴壁生；（２）培养４天，细胞开始生长呈梭形，胞核为椭圆形，有２～３个核仁；１０天后细胞融合成片生长，细胞间分界变得模糊不清；２０天后，部分细胞聚集成灶，胞体转变为立方形；５０天后的成骨细胞互相重叠生长，分泌的胶原纤维呈网状互相交织；（３）ＡＬＰ染色后的电镜观察显示，细胞膜的ＡｎＰ染色呈强阳性，而核膜、线粒体及内质网等细胞器及胞质的染色均为阴性。（４）胞体内富含线粒体和内质网。因此认为：成骨细胞在体外培养中仍具有旺盛的代谢能力，是体外成骨的前提。本研究为进一步研究人的骨代谢、生长调控和环境影响等提供了一个有益的体外实验模型。
AbstractHuman osteoblastic cells were obtained by explant-ing the femoral periosteum cultured in DMEM/F12medium and 15%fetal bovine serum.The variation oftheir apporance was obscrved in 50 days,and the elec-tron micrograph of osteoblasts stained by alkaline phos-phatase (ALP)on the 20th day,The result shows :(1)osteoblast is fibroblastic type cell.(2 ) osteoblast is spin-dle-shaped with 2～3 nucleolus in an oval nucleus aftercultu ring 4 days, Ten days after, osteoblasts contactwith each ot her and the memberane outline is blurred.The cells turned to be cube-shaped and gathered togeth-er in 20 days, then they overlapped and secreted colla-gen fibers which interwove each other in 50 days.(3)the elec tron micrograph showed that the membrane ofost eobl asts was stained strongly positive by ALP,butnegtive in cytoplasm, nuclear membrane, endoplasmicreticulum and mitochondrioa.(4)endoplasmic reticulumand mitochondrioa are rich in the endoplasm, The au-thors believe that osteoblasts metablize actively in vit-ro , and provide a possibility to form bone in vitro. Anexperimental medel for the study of bone mctabolism,growth control and environmental effects was estab-lished.
出处
《中华骨科杂志》
CAS
CSCD
北大核心
1995年第11期789-790,共2页
Chinese Journal of Orthopaedics
基金
国家自然科学基金
