Long-term maintenance of hematopoiesis in irradiated mice by retrovirally transduced peripheral blood stem cells
Long-term maintenance of hematopoiesis in irradiated mice by retrovirally transduced peripheral blood stem cells
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DOI：
10.1182/blood.v89.5.1811.1811_1811_1817
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发表时间：
1997-03-01
期刊：
影响因子：
20.3
通讯作者：
Zander, A
中科院分区：
文献类型：
--
作者：
Drize, N;Chertkov, J;Zander, A
动员后的外周血干细胞（PBSC）作为移植和基因治疗的造血干细胞来源，但其是否与正常骨髓造血干细胞完全等同，能否在重建小鼠体内提供长期的转基因功能，目前尚不清楚。用人腺苷脱氨酶基因（hADA）转导来自雄性小鼠的动员的PBSC，并用于重建致死辐射的雌性小鼠。在重建后1 12、3、6、9和12个月，在轻度麻醉下重复采集每只小鼠的骨髓细胞，通过聚合酶链反应（PCR）和Southern印迹杂交分析，研究CFUS来源的集落中脾集落形成单位（CFU-S）的数量、脾再生能力（SRA）和人ADA基因的整合。9个月后，在用动员的PBSC重建的小鼠中通过PCR用Y染色体特异性探针检测到的供体CFU-S的比例为75.3% +/-6.0%，这与骨髓移植后观察到的供体CFU-S的浓度相似。同样，在用转导的动员的PBSC或骨髓细胞重建的小鼠中，CFU-S的浓度没有差异。然而，在这两种情况下，骨髓中的CFU-S含量与移植非转导骨髓的小鼠中CFU-S的浓度相比降低了5倍至10倍。移植后1.5个月，骨髓细胞和外周血细胞重建的小鼠CFU-S的SRA相同，但4个月后，骨髓细胞重建的小鼠的SRA比PBSC移植的小鼠高5倍。在9个月期间，在约60%的研究CFU-S中观察到人ADA基因的整合。重组后1年，标记菌落的比例急剧下降。在整个观察期内，同一重组小鼠可同时出现1 ~ 9个单独标记的克隆，克隆存在时间约为3个月。我们的结论是，长期骨髓再生细胞动员到循环治疗粒细胞集落刺激因子（G-CSF）和干细胞因子（SCF）能够维持终身多克隆造血重建小鼠。(C)1997年，美国血液学会。
Mobilized peripheral blood stem cells (PBSC) are used as a source of hematopoietic stem cells for transplantation and gene therapy, It is still unclear, however, whether the PBSC are fully equivalent to normal bone marrow hematopoietic stem cells and whether they are able to provide long-term function of transgene in reconstituted mice, In the present study, mobilized PBSC from male mice were transduced with human adenosine desaminase gene (hADA) and were used for reconstitution of lethally irradiated female mice. At 1 1/2, 3, 6, 9, and 12 months after reconstitution, the bone marrow cells were repeatedly collected from each mouse under light anesthesia and the number of colony-forming unit-spleen (CFU-S), spleen repopulating ability (SRA), and the integration of human ADA gene were studied in CFUS-derived colonies by polymerase chain reaction (PCR) and Southern blot hybridization analyses. After 9 months, the proportion of donor CFU-S detected by PCR with a Y-chromosome-specific probe in mice reconstituted with mobilized PBSC was 75.3% +/- 6.0%, which is similar to the concentration of donor CFU-S seen after bone marrow transplantation. Similarly, there was no difference in the concentration of CFU-S in mice reconstituted with transduced mobilized PBSC or bone marrow cells. However, in both cases the CFU-S content in the bone marrow was reduced fivefold to 10-fold compared with the concentration of CFU-S in mice transplanted with nontransduced bone marrow. The SRA of CFU-S in mice reconstituted with peripheral blood and bone marrow cells was the same 1.5 months posttransplantation, but after an additional 4 months, SRA of mice reconstituted with bone marrow cells was fivefold higher as compared with those engrafted by PBSC. The integration of the human ADA gene was observed during 9 months in about 60% of studied CFU-S. The proportion of marked colonies sharply decreased 1 year following reconstitution. One to 9 individually labeled clones could be shown simultaneously by Southern blot hybridization in the same reconstituted mice during the whole period of observation, The time of clone existence was about 3 months. We conclude that long-term marrow repopulating cells mobilized into circulation by treatment with granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) are capable of maintaining lifelong polyclonal hematopoiesis in reconstituted mice. (C) 1997 by The American Society of Hematology.