4℃条件下保存不同时间点小鼠骨髓间充质干细胞的细胞活性和生物学特性研究  

Cellular Activity and Biological Characteristics of Mouse Bone Marrow Mesenchymal Stem Cells during Different Preservation Times at 4 ℃

作　　者：时炳正[1] 徐志媛[1] 杜晓岩[1] 范龙坤[2] 陈勇[2] 廖立[3] 金岩[3] 赵世俊 

机构地区：[1]佳木斯大学口腔医学院,黑龙江佳木斯154007 [2]沧州市中心医院口腔颌面外科 [3]第四军医大学口腔医学院组织工程中心 [4]沧州市人民医院口腔颌面外科

出　　处：《口腔医学研究》2013年第3期205-209,共5页Journal of Oral Science Research

基　　金：国家基础研究项目(973项目)(2010CB944800;2011CB964700);佳木斯大学重大项目培育基金项目(SZP2001-001);黑龙江省研究生创新科研项目(YJSCX2012-359HLJ)

摘　　要：目的:本研究旨在比较4℃条件下保存时间对小鼠骨髓间充质干细胞的细胞活性和生物学特性的影响,为安全有效短期保存BMSCs提供参考。方法:体外分离培养纯化小鼠骨髓间充质干细胞,使用磷酸盐缓冲液(Phosphate Buffered Saline PBS)为小鼠骨髓间充质干细胞的短期储存液,直接将骨髓间充质干细胞置于4℃保存,分别在0、4、12、24、48h观察细胞形态、活细胞比例、细胞增殖和分化能力。结果:小鼠骨髓间充质干细胞于4℃条件下保存0、4、12h和24h镜下见细胞形态良好、活细胞比例仍接近80%,细胞增殖和分化能力与0h相比未见显著性差异;保存至48h镜下见死细胞和细胞碎片增多、活细胞比例仅接近20%,细胞增殖和分化能力与0h相比存在显著性差异(P<0.05)。结论:4℃条件下24h内保存小鼠骨髓间充质干细胞,是一种简洁有效可行的短期保存方法。随保存时间的延长细胞逐渐丧失细胞活性和生物学特性。Objective： To compare the influence of cell activity and biological characteristics of mouse bone marrow mesenchymal stem cells （BMMSCs） at 4℃ during different preservation times, thus to provide a reference for safe and effective shortterm preservation of BMMSCs. Methods： After isolation, culture and purification, mouse BMMSCs were directly stored at 4℃ using phosphate- buffered saline （PBS） as short-term preservation solution. After Oh, 4h, 12h, 24h and 48h, morphology, living rate, proliferation and differentiation capacity of BMMSCs were detected. Results： After 0h, 4h, 12h and 24h of 4℃storage, the morphology of the mouse BMMSCs is nor- mal, with a living rate of 80%, while there was no significant differences in proliferation and differentiation capacities among these groups. When stored for 48h, an increasing number of dead cells and cell debris were observed in the medium of BMMSCs, the proportion of living cells declined to 20%, the proliferation and differentiation capaci- ties revealed significant differences compare with Oh （P〈0.05）. Conclusion： It is a simple and feasible method for short--term preservation of mouse BMMSCs at 4℃within 24h before further manipulation. But the activities and biological characteristics of BMMSCs would be lost gradually with the elongation of storage time.

关 键 词：骨髓间充质干细胞成骨诱导 成脂诱导 细胞悬液4℃保存 

分 类 号：R780.2[医药卫生—口腔医学]