Mature monocytic cells enter tissues and engraft
Mature monocytic cells enter tissues and engraft
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DOI：
10.1073/pnas.95.25.14944
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发表时间：
1998-12-08
影响因子：
11.1
通讯作者：
Abkowitz, JL
中科院分区：
文献类型：
--
作者：
Kennedy, DW;Abkowitz, JL
本研究的目的是鉴定作为组织巨噬细胞直接前体的循环细胞。ROSA 26骨髓单核细胞（含有编码β-半乳糖苷酶和新霉素抗性活性的β-geo转基因）在巨噬细胞集落刺激因子和flt 3配体存在下培养6天，以产生处于所有成熟阶段的单核细胞。将扩增的单核细胞（EMC）、未成熟（ER-MP 12（+））和更成熟（ER-MP20（+））亚群移植到辐照的B6/129 F2小鼠中。移植后15分钟，动物组织切片的β-半乳糖染色表明供体细胞随机着陆。到3小时，移植ER-MP20（+）（更成熟）EMC的动物中肺和肝脏中的供体细胞比移植未分离的EMC或新鲜骨髓单核细胞的动物中的供体细胞更频繁，这种模式持续到3天和7天。在第3天，在脾、肝、肺和脑（很少）中发现供体细胞作为簇以及单个细胞。到第7天和第14天，簇的大小增加，并且细胞表达巨噬细胞抗原F4/80，表明发生了进一步的复制和分化。使用新基因的PCR来定量来自移植动物的组织中的供体DNA的量，并证实ER-MP 20（+）EMC优先移植。这些数据证明成熟的单核细胞产生组织巨噬细胞。由于这些细胞可以在体外扩增和操作，它们可能是一个合适的目标群体的基因治疗溶酶体贮积病。
The goal of this study was to identify the circulating cell that is the immediate precursor of tissue macrophages. ROSA 26 marrow mononuclear cells (containing the beta-geo transgene that encodes beta-galactosidase and neomycin resistance activities) were cultured in the presence of macrophage colony-stimulating factor and flt3 Ligand for 6 days to generate monocytic cells at all stages of maturation. Expanded monocyte cells (EMC), the immature (ER-MP12(+)) and more mature (ER-MP20(+)) subpopulations, were transplanted into irradiated B6/129 F2 mice. beta-gal staining of tissue sections from animals 15 min after transplantation demonstrated that the donor cells landed randomly. By 3 h, donor cells in lung and liver were more frequent in animals transplanted with ER-MP20(+) (more mature) EMC than in animals transplanted with unseparated EMC or fresh marrow mononuclear cells, a pattern that persisted at 3 and 7 days. At 3 days, donor cells were found in spleen, liver, lung, and brain (rarely) as clusters as well as individual cells. By 7 and 14 days, the clusters had increased in size, and the cells expressed the macrophage antigen F4/80, suggesting that further replication and differentiation had occurred. PCR for the neogene was used to quantitate the amount of donor DNA in tissues from transplanted animals and confirmed that ER-MP20(+) EMC preferentially engrafted, These data demonstrate that a mature monocytic cell gives rise to tissue macrophages. Because these cells can be expanded and manipulated in vitro, they may be a suitable target population for gene therapy of lysosomal storage diseases.