长期培养的人脐带间充质干细胞PCNA、IL-6、IL-11和galectin-3的表达  

Expression of PCNA,IL-6,IL-11 and galectin-3 in human umbilical cord mesenchymal stem cells during long-term culture

作　　者：毛文哲[1] 许超[1] 李扬秋[2] 陈少华[2] 柳菁[3] 刘俊[1] 廖继东[1] 

机构地区：[1]暨南大学医学院田家炳医学实验中心,广东广州510632 [2]暨南大学医学院血液病研究所,广东广州510632 [3]暨南大学医学院生物化学教研室,广东广州510632

出　　处：《中国病理生理杂志》2012年第6期1051-1056,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81100353);中央高校基本科研业务费专项资金资助项目(No.21611447)

摘　　要：目的:探讨长期培养的人脐带间充质干细胞(hUC-MSCs)增殖细胞核抗原(PCNA)、白细胞介素-6(IL-6)、白细胞介素-11(IL-11)和半乳糖凝集素-3(galectin-3)mRNA及蛋白表达的变化情况,为hUC-MSCs的实验研究和临床应用提供实验资料和理论依据。方法:取剖宫产新生儿脐带,分离、传代培养hUC-MSCs;收集第3、8、18、28和33代细胞及培养上清,用qRT-PCR、ELISA及Western bloting检测PCNA、IL-6、IL-11和galectin-3 mRNA和蛋白水平。结果:(1)hUC-MSCs PCNA、IL-6、IL-11 mRNA及IL-6、IL-11蛋白的表达随培养传代次数增加而减少,第33代较第3代分别降低了33%、56%、37%和50.3%、58.9%,差异显著(均P<0.01)。(2)各代间galectin-3 mRNA表达无显著差异(P>0.05),且各代间蛋白表达亦无明显差异。结论:(1)体外长期传代培养过程中hUC-MSCs增殖能力和支持造血能力可能逐渐减弱甚至丧失。(2)体外长期传代培养可能对hUC-MSCs的免疫调节功能无显著影响,这有待进一步的实验验证。AIM： To investigate the changes of the mRNA and protein levels of proliferating cell nuclear antigen（PCNA）,interleukin-6（IL-6）,IL-11 and galectin-3 in human umbilical cord mesenchymal stem cells （hUC-MSCs） during long-term culture in vitro. METHODS： Human umbilical cords from neonates via caesarean section were collected, and the hUC-MSCs were isolated and sub-cultured. The cells and culture supernatants at passages 3, 8, 18, 28 and 33 were collected. The mRNA expression and protein secretion of PCNA,IL-6,IL-11 and galectin-3 in different passages of hUC-MSCs were detected by qRT-PCR, ELISA, and Western blotting. RESULTS： The mRNA expression of PCNA,IL-6 and IL-11 and protein secretion of IL-6 and IL-11 reduced gradually along with the passage.Comparing the 33rd passage with the 3rd passage of MSCs, the mRNA expression of PCNA,IL-6 and IL-11 decreased by 33%, 56% and 37%, and the protein secretion decreased by 50.3% and 58.9%, respectively. The differences were statistically significant （P〈0.01）. No statistically significant difference of galectin-3 mRNA expression and the protein band brightness （P〉0.05） in the MSCs among different passages was observed. CONCLUSION： The multiplication and haematogenesis support capabilities of the might decrease or even lose during long-term culture in vitro. Long-term culture might have no effect on hUC-MSCs immunoregulatory capability of hUC-MSCs, which requires further study.

关 键 词：人脐带间充质干细胞 长期培养 增殖细胞核抗原 白细胞介素6 白细胞介素11 半乳糖凝集素3 

分 类 号：R[医药卫生]