CAJ | 학술논문

目的观察体外长期培养人脐带间充质干细胞的生物活性变化特点及评估其安全性.方法在体外环境中对人脐带来源的间充质干细胞进行了长期的标准化培养,并延长了培养时间,对细胞的表面标记、端粒酶活性、细胞周期、凋亡率、核型及裸鼠成瘤性进行分析.结果常规体外培养下,间充质干细胞的表面标记持续稳定.端粒酶分析表明随着传代次数的增加,细胞的端粒酶活性表达逐渐降低,细胞周期正常未出现异常凋亡或癌变.核型分析表明细胞增殖过程中细胞核型无异常变化.裸鼠成瘤性实验表明细胞无成瘤性风险.结论间充质于细胞在体外标准化培养至第15代,细胞分化能力相对减弱.25代以后生长缓慢细胞生物学特征.培养到第30代,表型及基因型无明显变化,未发生恶性转变.
목적 관찰체외장기배양인제대간충질간세포적생물활성변화특점급평고기안전성.방법 재체외배경중대인제대래원적간충질간세포진행료장기적표준화배양,병연장료배양시간,대세포적표면표기、단립매활성、세포주기、조망솔、핵형급라서성류성진행분석.결과 상규체외배양하,간충질간세포적표면표기지속은정.단립매분석표명수착전대차수적증가,세포적단립매활성표체축점강저,세포주기정상미출현이상조망혹암변.핵형분석표명세포증식과정중세포핵형무이상변화.라서성류성실험표명세포무성류성풍험.결론 간충질우세포재체외표준화배양지제15대,세포분화능력상대감약.25대이후생장완만세포생물학특정.배양도제30대,표형급기인형무명현변화,미발생악성전변.
Objective To determine the alteration in biological features and safety profile of human umbilical cord- derived mesenchymal stem cells (hUC- MSCs) subjected to long- term culture in vitro.Methods A long-term standardized in vitro culture of hUC-MSCs was completed via prolonging the incubation period for analyses on cell surface markers,telomerase activity,cell cycle,incidence of apoptosis,karyotype,and tumorigenicity of nude mice.Results Standardized in vitro culture of hUC-MSCs resulted in stable expression of surface markers.This was associated with tapering,but not markedly increased,telomerase activity despite passages,as suggested by telomerase assessment.In line with absence of apoptotic anomaly or tumorigenesis in cell cycles,the inconsiderable variation in karyotype during cell proliferation did not yield potential risk of tumorigenicity in nude mice.Conclusion The hUC-MSCs cultured in vitro are associated with restrained differentiation capacity at the 15th passage,diminished proliferation potency at the 25th passage and absence of malignant transformation at the 30th passage,as indicated by insignificant alteration in phenotypes and genotypes.