成骨细胞超低温冻存前后生物学功能变化的研究  

Preliminary Study on Transformation of the Biological Function of the Cryopreserved Osteoblasts Cultured in vitro

作　　者：谈万业[1] 孙明霞[2] 魏奉才[1] 丁广耀 董作青[1] 刘云生[1] 

机构地区：[1]山东大学齐鲁医院口腔科,山东济南250012 [2]山东大学齐鲁医院整形外科,山东济南250012 [3]蓬莱市人民医院口腔科,山东蓬莱265600

出　　处：《华西口腔医学杂志》2006年第5期462-465,共4页West China Journal of Stomatology

基　　金：山东省自然科学基金资助项目(Y99C05)

摘　　要：目的观察超低温(液氮,-196℃)冻存对幼兔骨髓来源的成骨细胞体外培养的生长特性和生物学功能的影响。方法自幼兔骨髓获取基质干细胞,体外诱导分化并培养扩增,部分细胞进行液氮冻存,对冻存前后细胞的形态变化、增殖情况、克隆形成率、碱性磷酸酶(ALP)的合成能力、蛋白质合成能力及体外矿化能力进行对比观察,综合比较低温冻存对细胞功能的影响。结果冻存复苏细胞的增殖有一定的时间延后性,其大体形态与冻存前基本一致,超微结构略有变化;复苏细胞仍保持典型的成熟成骨细胞的特征,具有较强的合成ALP及蛋白质的能力,两组细胞无统计学差异(P>0.05);复苏细胞的体外矿化能力无明显变化。结论超低温保存成骨细胞对其生物学功能影响甚微,利用复苏细胞进行骨组织工程方面的研究是可行的。Objective To culture and amplify the young rabbit＇s bone marrow stromal cells （BMSCs） in vitro, and to observe the effect of hypothermia on the cells＇ growing behavior and biological function. Methods BMSCs were acquired from the rabbit＇ tibia bone marrow and induced to mature esteoblasts in vitro. The cultured cells growing well in vitro were preserved in liquid nitrogen. The anabiotic cells having cryopreserved for 1 week were chosen as the experimental group, and the routine 7th generation as the control group. Their biological function in comparion by the examination of morphological changes, cells＇ proliferation ability, colone forming ratio, synthesis ability of ALP and protein, mineralized nodes forming ability were observed. Results As contrast to the control groups, the anabiotic cells also grew and proliferated well in vitro except a little more slowly than before. They had the similar general shape in all the time segments, but a little differences in cells＇ ultrastructure. The experimental groups also had the typical characters＇of mature esteoblasts, and high abilities of the synthesis of ALP and proteins. The statistic data showed that these two groups had no significant difference （P〉0.05）. Conclusion The cryopreserved osteoblasts had the same biological functions and the similar growing behaviors as before. These results suggest that it is practical to use the cryopreserved esteoblasts for further study on bone tissue engineering.

关 键 词：骨髓基质干细胞 成骨细胞 细胞培养 低温冻存 骨组织工程 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R318.52[医药卫生—基础医学]