骨髓基质细胞分泌物促进PC12细胞的分化和存活  

Secretory factors of bone marrow stromal cells promoting differentiation and survival of PC12 cells

作　　者：郑梅[1] 樊东升[1] 张俊[1] 宋德懋[2] 范少光[2] 

机构地区：[1]北京大学第三医院神经科 [2]北京大学第三医院基础医学院生理与病理生理学系

出　　处：《中华神经医学杂志》2005年第1期20-23,34,共5页Chinese Journal of Neuromedicine

基　　金：国家高技术研究发展计划(863计划)基金资助项目(2001AA217161)

摘　　要：目的体外检测骨髓基质细胞分泌物对PC12细胞的活性作用,并探讨其产生的可能机制。方法收集培养至第4代第7天的SD大鼠MSCs培养上清,按不同的体积百分比浓度加入到PC12细胞培养体系中,在倒置相差显微镜下观察1 d和4 d的细胞形态学改变;用丙二酸钠对PC12细胞造成氧化应激损伤,同时加入不同体积百分比浓度的MSCs培养上清,采用MTT法测定24 h后的细胞活性。结果有突细胞/总细胞数、最长突起长度随培养时间及MSC培养上清体积百分比的增加而增加;PC12细胞氧化损伤后,加入一定浓度MSC培养上清组的PC12细胞活性较未加入组增高,差异有显著性。结论MSCs能够合成和分泌具有神经营养活性的物质,该物质能诱导PC12细胞分化并减轻氧化应激对PC12细胞的损伤。Objective To investigate the effects of bone marrow stromal cells (BMSCs) secretory factors on PC12 cells cultured in vitro and to discuss its probable mechanism. Methods The BMSCs of SD rats were cultured to the 4th generation and the supernatant on the 7th day was collected and added to the cultured PC12 cells at different volume percentage concentrations. The morphology of PC12 cells was observed using phase-contrast microscope on day 1 and 4. Meanwhile, after treatment with malonate to result in a damage of oxidative stress to the PC12 cells, supernatant of BMSCs at different volume percentage concentrations was added to the damaged PC12 cells, followed by examining the cyto-viability using MTT method after 24 hours. Results The elevation of process length and ratio of cells with process to cells in gross number was paralleled to the rise of concentrations of BMSCs supernatant and culture period (Ftime=124.78, P<0.0001; Fconcentration=117.8, P<0.0001). The cyto-viability of damaged PC12 cells with a certain concentration of MSCs supernatant was enhanced compared with that without MSCs supernatant (Ftime=56.68, P< 0.0001; Fconcentration=22.33, P< 0.0001). Conclusion BMSCs can synthesize and secrete neurotrophic factors physiologically, which may induce PC12 cells differentiation and protect PC12 cells from damage of oxidative stress.

关 键 词：PC12细胞 培养上清 骨髓基质细胞 分泌物 分化 存活 MSC 神经营养活性 氧化应激损伤 培养体系 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]