人脐带间充质干细胞对脐血CD34~+细胞在NOD/SCID小鼠体内造血重建的影响

目的 探讨人脐带间充质干细胞对脐血CD34~+细胞在NOD/SCID小鼠体内造血重建的影响.方法 将3.5×10~5个脐血CD34~+细胞单独(单移植组)或与5.0×10~6个人脐带间充质干细胞共同(共移植组)输入经~(137)Cs 3.0Gy照射后的NOD/SCID小鼠体内,观察移植后6周内小鼠外周血象的变化情况.于移植后第6周处死小鼠,采用流式细胞术检测小鼠骨髓、脾脏及外周血人源细胞(hCD45~+)含量,并分别检测小鼠骨髓中人源淋巴系(CD3/CD19)、粒系(CD33)、单核系(CD14)、血小板(CD61)、红系(CD235a)等各系血细胞比例,比较间充质干细胞共移植对CD34~+细胞植入率的影响.结果 移植后3周,两组小鼠外周血象开始有不同程度恢复;移植后6周,共移植组外周血白细胞和血小板计数均已达高峰,明显高于单移植组(P<0.05),两组小鼠的红细胞计数差异无统计学意义(P>0.05).移植后6周,共移植组骨髓及外周血中人源细胞hCD45~+CD34~+比例分别为(42.66±2.57)%和(4.74±1.02)%,明显高于单移植组的(25.27±1.67)%和(1.19±0.54)%(P=0.006).移植后6周,共移植组小鼠骨髓内的CD19~+、CD33~+、CD14~+、CD61~+和CD235a~+细胞比例均明显高于单移植组(P<0.05),CD3~+T淋巴细胞比例明显低于单移植组(P=0.003);CD19~+B淋巴细胞得到优势扩增,明显高于其他各系血细胞比例(P<0.05).结论 脐带间充质干细胞与脐血CD34~+细胞共移植可促进造血干细胞的植入,缩短CD34~+细胞移植后造血恢复时间.

Effect of Human Umbilical Cord Mesenchymal Stem Cells on the CD34~+ Cells Transplantation in NOD/SCID Mice

Objective To study the effect of human umbilical blood(UB)mesenchymal stem cells(MSC)on the CD34+ cells transplantation in NOD/SCID Mice.Methods Umbilical blood CD34~+ cells(3.5×10~5 cells)alone or combined with umbilical cord MSC cells were transplanted into NOD/SCID mice that had been irradiated with~(137) Cs(3.0Gy)before transplantation.Changes in peripheral blood cells within 6 posttransplantation weeks were detected.The mice were sacrificed 6 weeks after transplantation.The human hematopoietic cells(hCD45~+)and multi-lineage engraftment cells(CD3/CD19,CD33,CD14,CD61,and CD235a) in NOD/SCID recipients' bone marrow,spleen,and peripheral blood were analyzed by flow cytometry.Results In the 3rd post-transplantation week,white blood cells(WBC),platelets(PLT),and red blood cells(RBC)began to increase in both two groups.In the 6th post-transplantation week,WBC and PLT counts in CD34~+ +MSC group reached peak levels and were significantly higher than CD34~+ alone group(P <0.05),while RBC level was not significantly different between these two groups(P > 0.05).hCD45~+ cell levels in bone marrow and peripheral blood were(42.66±2.57)% and(4.74±1.02)% in CD34~+ +MSC group,which were significantly higher than those in CD34~+alone group(25.27±1.67)%and(1.19±0.54)%,respectively,P = 0.006].Also in the 6th post-transplantation week,the proportions of CD19~+,CD33~+,CD14~+,CD61~+,and CD235a~+in CD34~++MSC group were significantly higher than those in CD34~+ alone group(P < 0.05),while the proportion of CD3~+ T lymphocyte in CD34~+ +MSC group was significantly lower than that in CD34~+alone group(P =0.003).The amplification of CD19~+ Blymphocyte was significantly higher than other blood cell lingeages(P < 0.05).Conclusion The co-transplantation of MSC cells and CD34~+cells can promote hematopoetic stem cell transplantation and hematopoetic recovery in vivo.