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• Jerebtsova, M
• Kumari, N
• Xu, M
• Melo, G
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• Jeang, K
• Nekhai, S
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• Kumari, N
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Biology 2012, 1(2), 175-195; doi:10.3390/biology1020175

Article

HIV-1 Resistant CDK2-Knockdown Macrophage-Like Cells Generated from 293T Cell-Derived Human Induced Pluripotent Stem Cells

Marina Jerebtsova ^1,2 , Namita Kumari ³ , Min Xu ³ , Gustavo Brito Alvim de Melo ⁴ , Xiaomei Niu ³ , Kuan-Teh Jeang ⁴  and Sergei Nekhai ^3,5,*

¹ Center for Molecular Physiology Research, Children’s National Medical Center, Washington, DC 20010, USA ² Department of Pediatrics, the George Washington University School of Medicine and Health Sciences, Washington, DC 20037, USA ³ Center for Sickle Cell Disease, Department of Medicine, Howard University, Washington, DC 20059, USA ⁴ Molecular Virology Section, Laboratory of Molecular Microbiology, NIAID, National Institutes of Health, Bethesda, MD 20892, USA ⁵ 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

(This article belongs to the Special Issue Structural and Molecular Biology of HIV)

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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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