利用新生大鼠建立人/大鼠造血嵌合体模型

Development of the Human/Rat Chimera Model with Neonatal Rats

本实验旨在通过输注脐血Lin-细胞 ,利用新生大鼠建立人 /大鼠造血嵌合体模型。将脐血来源的Lin-细胞移植到正常一级新生SD大鼠的肝脏内 ,移植后的新生大鼠与同窝未接种的正常新生大鼠共同饲养和观察 ,于输注Lin-细胞后 2 ,4和 8周时取嵌合鼠的外周血、脾脏等组织 ,检测人源细胞的比例及人特异性 β2 微球蛋白基因片段。结果发现 ,通过流式细胞术检测到嵌合鼠外周血中存在有一定比例的人源细胞 ,PCR证明了嵌合鼠脾脏基因组DNA中存在人特异性 β2 微球蛋白基因片段。结论 :利用新生大鼠可以建立人造血嵌合体 ,它为今后利用此实验模型进行人造血干细胞体内生物学特性的检测及其它实验奠定了基础 。

The purpose of this study was to transplant neonatal rat with human cord blood Lin - cells to test the possibility of this xenograft model. The Lin - cells were purified from human cord blood (CB) using negative selection strategy based on different lineage specific antigens. The Lin - cells were injected into the liver of neonatal rats using a microinjector at an average of 5×10 5 cells for each. Peripheral blood (PB) and spleen were collected at 2,4 and 8 weeks after injection. Flow cytometry was performed to detect human cells in the rat PB, PCR was used to detect human cells in PB as well as spleen. The results showed that a definite proportion of human cells existed in peripheral blood of chimeric rat and the human specific β2 microglobulin gene fragments were detected in spleen genomic DNA of chimeric rat. It is concluded that human /rat chimera model can be developed with neonatal rats. Human/rat xenograft model may provide a useful and convenient method for human hematopoietic stem cell assay in vivo.