CAJ | 학술논문

目的探讨骨髓间充质干细胞(MSC)对脐血造血干细胞体外扩增及抗细胞衰老的作用.方法从新鲜脐血中分离出CD34+细胞,分别接种在含或不含MSC及细胞因子的培养体系中,取培养10d后的造血干细胞,分别用于细胞计数,集落分析,流式细胞仪检测表面标记,并对培养后的细胞进行β-半乳糖苷酶活性检测,采用实时定量聚合酶链反应(PCR)检测衰老相关基因p16INK4amRNA的表达.结果体外培养10d后,MSC组、细胞因子组及MSC+细胞因子组对脐血总有核细胞(TNC)、CD34+细胞的体外扩增及克隆形成能力均有支持作用,以MSC+细胞因子组最为明显(P＜0.05),其扩增倍数分别达到(45.3±6.8)倍、(38.4±5.8)倍及(50.2±4.2)倍,但衰老细胞比例及p16INK4amRNA表达倍数则以MSC组最低,MSC+细胞因子组次之,细胞因子组两项指标均为最高(P＜0.05).结论 MSC能更好保护体外扩增的脐血造血干细胞,减少细胞衰老的发生,MSC+细胞因子是有效扩增脐血造血干细胞并保持细胞干性的更理想方法.
목적 탐토골수간충질간세포(MSC)대제혈조혈간세포체외확증급항세포쇠로적작용.방법 종신선제혈중분리출CD34+세포,분별접충재함혹불함MSC급세포인자적배양체계중,취배양10d후적조혈간세포,분별용우세포계수,집락분석,류식세포의검측표면표기,병대배양후적세포진행β-반유당감매활성검측,채용실시정량취합매련반응(PCR)검측쇠로상관기인p16INK4amRNA적표체.결과 체외배양10d후,MSC조、세포인자조급MSC+세포인자조대제혈총유핵세포(TNC)、CD34+세포적체외확증급극륭형성능력균유지지작용,이MSC+세포인자조최위명현(P＜0.05),기확증배수분별체도(45.3±6.8)배、(38.4±5.8)배급(50.2±4.2)배,단쇠로세포비례급p16INK4amRNA표체배수칙이MSC조최저,MSC+세포인자조차지,세포인자조량항지표균위최고(P＜0.05).결론 MSC능경호보호체외확증적제혈조혈간세포,감소세포쇠로적발생,MSC+세포인자시유효확증제혈조혈간세포병보지세포간성적경이상방법.
Objective To investigate the effects and mechanisms of bone marrow mesenchymal stem cells (MSCs) on cord blood hematopoietic cells after culture ex vivo with or without cytokines,and and develop an optimal culture system for cord blood hematopoietic cells culture ex vivo.Method CD34+ cells separated from cord blood were cultured for 10 days.Total nucleated cells (TNCs),CD34+ cells and colony-forming unit (CFU) were analyzed.SA-β-gal and p16 mRNA was detected by using real-time reverse transcriptase (PCR).Results Ex vivo results demonstrated that the three culture systems supported expansion of TNCs,CD34+ cells and CFU,especially in the MSCs + cytokine group.The proportion of senescent cells and p16 mRNA expression in the expanded cells in the MSCs group were lower than those in the other two groups (P＜0.05).Conclusion MSCs are capable of protecting hematopoietic cells and reduce the occurrence of cell senescence in expanded cells.MSCs in combination with cytokines could efficiently support proliferation of cord blood hematopoietic cells cultured ex vivo.