Forced expression of human macrophage colony-stimulating factor in CD34+ cells promotes monocyte differentiation in vitro and in vivo but blunts osteoclastogenesis in vitro
Forced expression of human macrophage colony-stimulating factor in CD34+ cells promotes monocyte differentiation in vitro and in vivo but blunts osteoclastogenesis in vitro
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DOI：
10.1111/ejh.12867
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发表时间：
2017-05-01
影响因子：
3.1
通讯作者：
Richter, Johan
中科院分区：
文献类型：
--
作者：
Almendras, Carmen P. Montano;Thudium, Christian S.;Richter, Johan
目的在体外和体内验证人M-CSF （hM-CSF）在脐带血（CB） CD34（+）细胞中过表达诱导单核细胞和破骨细胞分化和存活的假设。方法利用慢病毒载体在脐带血CD34（+）细胞中过表达人M-CSF。结果我们发现，与对照细胞相比，lv - hm - csf转导的CB CD34（+）细胞在暴露于表达hm - csf的载体1次或2次后分别扩增3.6倍和8.5倍。同样，lv - hm - csf转导的CB CD34（+）细胞显示出明显更高的单核细胞水平。此外，这些细胞产生高水平的hM-CSF。此外，它们能够在体外分化为功能性骨吸收破骨细胞。然而，与接受外源性hM-CSF的对照CD34（+）细胞相比，破骨细胞的分化和骨吸收受到抑制。移植了lv -hM-CSF转导的CB CD34（+）细胞的NSG小鼠产生了生理水平的hM-CSF，导致外周血、骨髓以及脾、肺和肝脏中人单核细胞的百分比增加。总之，在体外和体内，人CD34（+）细胞中的M-CSF异位产生促进细胞扩增和单核细胞分化，并允许形成功能性破骨细胞，尽管在没有外源M-CSF的情况下，体外水平降低。
ObjectivesHere, we tested the hypothesis that human M-CSF (hM-CSF) overexpressed in cord blood (CB) CD34(+) cells would induce differentiation and survival of monocytes and osteoclasts in vitro and in vivo.MethodsHuman M-CSF was overexpressed in cord blood CD34(+) cells using a lentiviral vector.ResultsWe show that LV-hM-CSF-transduced CB CD34(+) cells expand 3.6- and 8.5-fold more with one or two exposures to the hM-CSF-expressing vector, respectively, when compared to control cells. Likewise, LV-hM-CSF-transduced CB CD34(+) cells show significantly higher levels of monocytes. In addition, these cells produced high levels of hM-CSF. Furthermore, they are able to differentiate into functional bone-resorbing osteoclasts in vitro. However, osteoclast differentiation and bone resorption were blunted compared to control CD34(+) cells receiving exogenous hM-CSF. NSG mice engrafted with LV-hM-CSF-transduced CB CD34(+) cells have physiological levels of hM-CSF production that result in an increase in the percentage of human monocytes in peripheral blood and bone marrow as well as in the spleen, lung and liver.ConclusionIn summary, ectopic production of human M-CSF in CD34(+) cells promotes cellular expansion and monocyte differentiation in vitro and in vivo and allows for the formation of functional osteoclasts, albeit at reduced levels, without an exogenous source of M-CSF, in vitro.