摘要
目的 :观察大鼠骨组织来源成骨细胞的体外分离培养的条件及生长特征 ,为骨组织工程学实验研究奠定基础。方法 :取出生 2 4hSD乳鼠的顶骨 ,酶消化法分离培养成骨细胞并传代 ,观察其生物学特征 ,并进行ALP活性测定 ,绘制生长曲线、分裂指数曲线和贴壁率曲线 ,测定细胞贴壁及延展时间。并将培养细胞进行冻存、复苏 ,比较细胞特性有无改变。结果 :培养细胞在第 8代以前生长性状稳定 ,具有典型的成骨细胞特性 ,可作为实验细胞 ;第 4d为细胞生长倍增时间 ,第 5～ 6d细胞达生长高峰 ;第 4d细胞分裂最为旺盛 ,达 18% ;传代后 10h贴壁率最高 ,为 90 % ;冻存复苏后的细胞生物学特性无改变。结论 :本实验方法体外培养的成骨细胞 ,生长性状稳定 ,增殖速度快 ,具有与体内成骨细胞相同的生物学特性 ,保证了相关实验的可靠性和准确性 。
Objective: To observe culture condition and growth characteristics of osteoblasts in vitro ,which were developed from bone tissue of mice,contributing to the bone tissue engineering experiment.Methods:Osteoblasts,which were developed from parietal bones of SD mice with a mean age of twenty four hours after enzyme isolation,were cultured and subcultured to investigate their biological features,to evaluate ALP activity,to assess their strapping and expanding time and to draw graphs of growth,split index and adhesion rate.Frozen and resuscitated cells,after culturing,were compared to understand the differences of their features.Result:Features of cultured cells,which can be used as experiment cells,with typical features of osteoblasts,were steady before the eighth passage;Doubling time of cells was the fourth day,the highest proliferation was on the 5 th 6 th day;mitosis index of the cells was the highest on the fourth day (18%);Adhesion rate of the cells after passage of ten hours was the highest(90%).The biological features of cells have no changes after freezing and resuscitation.Conclusion:Osteoblasts cultured in vitro by this method,characterized of steady growth,rapid proliferation and the same biological features with osteoblasts in vivo,are proved to assure experiment of authenticity and accuracy and to be applied to the study of bone tissue engineering as seed cells.
出处
《中国矫形外科杂志》
CAS
CSCD
2002年第5期470-472,共3页
Orthopedic Journal of China
