成骨细胞的体外培养及其生物学特性

目的 :建立体外分离、培养兔和人成骨细胞的生物学模型 ,并对其生物学特性进行观察。方法 :抽取新西兰兔骨髓 ,采用全骨髓法培养 ;人髂骨松质骨经胰酶消化获得成骨细胞 ,以 1× 10 6 / ml的细胞浓度进行培养 ,约10 d左右得到贴壁生长的单层细胞。随后传代培养 ,对所获得的细胞进行生物学特性观察。 结果 :获得的细胞呈多种形态 ,有长短不一、粗细不均、互相连接的胞浆突起 ,细胞可相互重叠呈复层生长 ,并形成钙结节。经连续传代 ,细胞形态与功能不变 ,具有典型成骨细胞的生物学特性。结论:无论采用兔骨髓基质细胞全骨髓法 ,还是人松质骨经胰酶消化法均能在体外培养出大量高纯度成骨细胞 ,方法简便、易行 ,为成骨细胞复合生物降解材料移植修复骨缺损奠定了基础 ,而且培养的成骨细胞可作为生物学模型 ,供干预研究使用。

Biologic features of osteoblast and its culture in vitro

Objective: To cultivate the osteoblast cells of rabbit and human in vitro and study its biological characteristics. Methods: To obtain the stromal cells of bone marrow from the trochanteric region of femur of rabbit and osteoblast from cancellous bone of normal human with enzyme digestion method and the cells were cultured with special cultural fluid in vitro. After about ten days,cells lined up in one layer. Then acquired cells were generationed and identified with inverted microscope observation,HE stains,alkaline phosphorase stain,Von Kossa stain of calcified nodule. Result: The cells had the same morphological feature with the classical osteoblast cells. ALP activity,Von Kossa stain of calcified nodule were both positive. Conclusions: It is easy to purify and culture plentiful osteoblasts with normal functions not only by enzyme digesting cancellous bone of normal human but also the stromal cells of bone marrow of rabbit. The cultured osteoblast in vitro were a successful model as intervention study.