摘要
目的研究脉冲电磁场对免疫磁性分选人前软骨干细胞（PSCs）增殖功能的影响。方法采用酶消化法从流产胎儿下肢股骨干骺端中收集细胞，选用免疫磁性分选技术分离出PSCs；经体外扩增培养后采用流式细胞术、免疫组化、免疫荧光、RT—PCR等技术鉴定纯化后的PSCs。采用50Hz 1mT脉冲电磁场间断刺激纯化后的PSCs，并绘制细胞生长曲线，采用MTT法测定细胞增殖活性，采用流式细胞术检测细胞周期及细胞凋亡情况。结果从流产胎儿干骺端中成功培养出细胞表面表达成纤维生长因子受体-3（FGFR-3）标记物的人PSCs；免疫磁性分选系统能有效分离纯化PSCs；PSCs经脉冲电磁场刺激4d和6d后，发现能明显促进细胞增殖功能（P〈0．05），其S期细胞数量百分比较对照组显著增高（P〈0．01），细胞早期凋亡率、晚期凋亡率及总凋亡率均较对照组明显降低（P〈0．05）。结论免疫磁性分选系统能有效分离、纯化PSCs，脉冲电磁场刺激能显著提高体外培养人PSCs增殖功能并抑制细胞凋亡。
Objective To study the biological effects of pulsed electromagnetic fields （PEMFs） on the proliferation of immunomagnetically separated human precartilaginous stem cells （PSCs） in vitro. Methods The cells from an aborted fetus＇s metaphysis were digested using collagenase. The PSCs were isolated by magnetic cell sorting （MACS）, then subcultured and amplified. Flow cytometry, immunohistochemistry, immunofluorescence and RTPCR analysis were performed to identify the purified PSCs. The PSCs were stimulated by PEMFs at 50 Hz frequency and 1 mT intensity. Cell proliferation was measured at different time points using methyl thiazolyl tetrazolium （MTT） , and the cell growth curve was plotted. Flow cytometry was applied to measure the cell cycle and apoptosis. Results The PSCs were successfully cultured. There was fibroblast growth factor receptor-3 （FGFR-3） on their surface. Cell proliferation was promoted after 4 and 6 days of PEMF stimulation. The percentage of cells at the S phase was higher than in a control group. Early, late and total rates of apoptosis in the experimental group decreased significantly. Conclusion PEMFs can enhance the proliferation and inhibit the apoptosis of human PSCs, and it is possible to cultivate the high density human PSCs in vitro.
出处
《中华物理医学与康复杂志》
CAS
CSCD
北大核心
2009年第5期296-300,共5页
Chinese Journal of Physical Medicine and Rehabilitation
基金
国家自然科学基金专项基金项目（30650007）
关键词
前软骨干细胞
成纤维生长因子受体-3
细胞培养
脉冲电磁场
Precartilaginous stem cells
Fibroblast growth factor receptor-3
Cell culture
Pulsed electromagnetic fields
