摘要
目的：探讨低氧处理对不同年龄C57小鼠骨髓间充质干细胞（BMSCs）增殖的影响。方法：全骨髓法从3~6周年轻C57小鼠（年轻组）股骨和胫骨的骨髓中提取、分离BMSCs,传代至第6代,部分于37℃、5%CO2条件下中常规培养,连续5d观察细胞的形态及数目,绘制细胞生长曲线;部分细胞进行成骨和成脂诱导。另取第6代BMSCs,随机分为低氧处理组（37℃、5%O2条件下培养30min,再常规培养）和阴性对照组（不行低氧处理,仅常规培养）,常规培养8h后固定细胞,结晶紫染色。观察两组细胞处理前后的数量变化。全骨髓法提取18~24个月老龄C57小鼠（老龄组）的骨髓细胞常规培养,连续12d观察细胞的形态及数目并绘制细胞生长曲线。将老龄组C57小鼠的骨髓贴壁细胞分为低氧处理组和阴性对照组,处理方法同年轻小鼠低氧处理和阴性对照组,于常规培养24h后固定细胞,结晶紫染色,观察两组细胞处理前后的数量变化。结果：年轻C57小鼠BMSCs增殖能力强,可传代至第6代,经过5d的培养细胞数量逐渐增加,并具有成骨、成脂诱导分化能力;但经低氧处理后,BMSCs呈片状脱落。老龄C57小鼠的骨髓贴壁细胞增殖能力差,经过12d的原代培养,细胞数量逐渐减少,并且只有一个集落形成;经低氧处理,贴壁细胞数量锐减,无新的集落形成。结论：低氧处理不能在体外扩增阶段提高C57小鼠BMSCs的增殖能力。
Objective：To investigate the effects of hypoxia on the proliferation of bone marrow mesenchymal stem cells（BMSCs）isolated from C57 mice of different age.Methods：BMSCs were isolated from the marrow of femurs and tibias of young C57 mice aged 3to 6weeks（young group）by whole bone marrow adherence method,and they were cultured.The 6th generation of BMSCs were routinely cultured under the condition of 5% CO2and37 ℃,their potential of differentiation to osteocytes and adipocytes were observed,and the cell numbers were counted,and a growth curve of 5consecutive days was drawn.Another portion of the 6th generation of BMSCs was randomly divided into hypoxia group and non-hypoxia group.The cells of hypoxia group were cultured under the condition of 5% O2 and 37 ℃for 30 minutes,followed by routine culture for 8hours,while the non-hypoxia group was routinely cultured continously.The change in the cell number of the two groups was observed with crystal violet staining.Adherent cells were isolated from old C57 mice aged 18 to 24months（aging group）with whole bone marrow adherence method.The cells were routinely cultured,the number of cells were observed and a growth curve of 12 consecutive days was drawn.The bone marrow adherent cells of old C57 mice were divided into two groups,and they were treated in same way as the young hypoxia group.After a routine culture of 24 hours,the change in the cell number was observed with crystal violet staining.Results：The proliferation ability of BMSCs of young C57 mice was robust,and they could be sustainable up to the 6th generation.The number of cells gradually grew in 5days,and they showed the potential of osteogenic or adipogenic differentiation.However,the cells fell off in flakes under hypoxia treatment.The proliferation ability of bone marrow adherent cells of aging C57 mice was poor.After a primary culture of 12 days,the cell number was gradually decreased,and only one colony was found to be formed.The number of adherent cells decreased obviously under the hypoxi
出处
《感染．炎症．修复》
2015年第1期23-27,F0002,共6页
Infection Inflammation Repair
基金
国家重点基础研究发展计划(973计划)项目(2012CB518105)
