第三方脐血细胞辅助输注对小鼠单倍型造血干细胞移植后造血重建的影响

The influence of umbilical cord blood cells infusion on hematopoietic reconstitution in a haploidentical hematopoietic stem cell transplantation mouse model

目的分析以脐血细胞作为第三方细胞辅助输注对小鼠单倍型造血干细胞移植后造血重建的影响。方法以CB6/F1雄性小鼠为供鼠,以BALB/C小鼠为受鼠。受鼠经Co60全身照射清髓后回输供鼠干细胞,建立单倍型造血干细胞移植模型。将40只受鼠分为对照组和实验组,每组各20只,对照组受鼠回输供鼠干细胞,实验组受鼠回输供鼠干细胞+脐血单个核细胞。于移植后+7d、+14d、+21d、+28d和+50d对存活受鼠进行断尾采血,检测白细胞、血红蛋白和血小板,分析造血重建情况。处死小鼠时取外周血1mL,采用荧光原位杂交技术检测Y染色体比例,计算嵌合率。两组受鼠各时间点白细胞、血红蛋白及血小板检测结果采用成组t检验进行比较,嵌合率采用卡方检验进行比较。P<0.05为差异有统计学意义。结果移植后+14 d实验组和对照组白细胞数量分别为(8.4±2.6)×10^9/L和(3.8±1.0)×10^9/L,差异有统计学意义(t=6.968,P<0.05);其他时间点两组白细胞数量差异无统计学意义。移植后两组受鼠各时间点血红蛋白检测结果差异均无统计学意义。移植后+7 d实验组和对照组血小板数量分别为(125±40)×10^9/L和(64±15)×10^9/L,移植后+14 d分别为(282±47)×10^9/L和(163±41)×10^9/L,差异均有统计学意义(t=6.366和8.093,P均<0.05);其他时间点两组血小板数量差异无统计学意义。结论使用脐血细胞作为第三方细胞辅助输注后,可促进单倍型造血干细胞移植受鼠早期白细胞和血小板较快恢复。

Objective To analyze the hematopoietic reconstitution in the mouse model of haploidentical hematopoietic stem cell transplantation when using umbilical cord blood cells as the third party cells.Methods The CB6/F1 mice were selected as the donor and the BALB/C mice were selected as the receptor.All the recipient mice were infused with donor stem cells after myeloablative with Co60 to establish the mouse model of the haploidentical hematopoietic stem cell transplantation.Forty recipient mice were equally divided into the experimental group and the control group.The mice in the control group were infused with donor mice-derrived haploidentical hematopoietic stem cell,and the mice in the experimental group were infused with umbilical cord blood cells and donor mice-derrived haploidentical hematopoietic stem cell.Blood collection by tail snipping in the viable recipient mice was performed at intervals of+7,+14,+21,+28,+50 days after transplantation and then the assay of white blood cell,hemoglobin and blood platelet were performed to evaluate the hematopoietic reconstitution.One mL peripheral blood was obtained when the mice were sacrificed.The proportion of Y chromosome was dected by fluorescence in situ hybridization and then the proportion of chimerism was calculated.The test results of white blood cell,hemoglobin and blood platelet between the 2 groups were compared by t test,and the proportion of chimerism were compared by chi-square test.P<0.05 was considered statistically significant.Results The white blood cell counts at+14 days after transplantation between the experimental group and the control group were(8.4±2.6)×10^9/L and(3.8±1.0)×10^9/L respectively,which had statistical significance(t=6.968,P<0.05),while no statistical signicance was found at other time points.No statistical significance was found between the 2 groups for hemoglobin.The blood platelet counts at+7d,+14d after transplantation between the experimental group and the control group were(125±40)×10^9/L and(64±15)×10^9/L,(282±47)×10^9/L and(163±41)×10^9/L respectively,which had statistical significance(t=6.366 and 8.093,P<0.05),while no statistical signicance was found at other time points.Conclusions The recovery of peripheral blood white blood cell and blood platelet in the mouse model of haploidentical hematopoietic stem cell transplantation was faster with the help of umbilical cord blood cells infusion.