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HIV-1 Resistant CDK2-Knockdown Macrophage-Like Cells Generated from 293T Cell-Derived Human Induced Pluripotent Stem Cells
Center for Molecular Physiology Research, Children’s National Medical Center, Washington, DC 20010, USA
Department of Pediatrics, the George Washington University School of Medicine and Health Sciences, Washington, DC 20037, USA
Center for Sickle Cell Disease, Department of Medicine, Howard University, Washington, DC 20059, USA
Molecular Virology Section, Laboratory of Molecular Microbiology, NIAID, National Institutes of Health, Bethesda, MD 20892, USA
Department of Microbiology, Howard University, Washington, DC 20059, USA
* Author to whom correspondence should be addressed.
Received: 15 May 2012; in revised form: 11 July 2012 / Accepted: 16 July 2012 / Published: 26 July 2012
Abstract: A major challenge in studies of human diseases involving macrophages is low yield and heterogeneity of the primary cells and limited ability of these cells for transfections and genetic manipulations. To address this issue, we developed a simple and efficient three steps method for somatic 293T cells reprogramming into monocytes and macrophage-like cells. First, 293T cells were reprogrammed into induced pluripotent stem cells (iPSCs) through a transfection-mediated expression of two factors, Oct-4 and Sox2, resulting in a high yield of iPSC. Second, the obtained iPSC were differentiated into monocytes using IL-3 and M-CSF treatment. And third, monocytes were differentiated into macrophage-like cells in the presence of M-CSF. As an example, we developed HIV-1-resistant macrophage-like cells from 293T cells with knockdown of CDK2, a factor critical for HIV-1 transcription. Our study provides a proof-of-principle approach that can be used to study the role of host cell factors in HIV-1 infection of human macrophages.
Keywords: HIV-1 resistant macrophage-like cells; CDK2 knockdown; iPSC
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Jerebtsova, M.; Kumari, N.; Xu, M.; Melo, G.B.A.; Niu, X.; Jeang, K.-T.; Nekhai, S. HIV-1 Resistant CDK2-Knockdown Macrophage-Like Cells Generated from 293T Cell-Derived Human Induced Pluripotent Stem Cells. Biology 2012, 1, 175-195.
Jerebtsova M, Kumari N, Xu M, Melo GBA, Niu X, Jeang K-T, Nekhai S. HIV-1 Resistant CDK2-Knockdown Macrophage-Like Cells Generated from 293T Cell-Derived Human Induced Pluripotent Stem Cells. Biology. 2012; 1(2):175-195.
Jerebtsova, Marina; Kumari, Namita; Xu, Min; Melo, Gustavo Brito Alvim de; Niu, Xiaomei; Jeang, Kuan-Teh; Nekhai, Sergei. 2012. "HIV-1 Resistant CDK2-Knockdown Macrophage-Like Cells Generated from 293T Cell-Derived Human Induced Pluripotent Stem Cells." Biology 1, no. 2: 175-195.