Generation of Monocyte-Derived Macrophages

Monocytes-derived macrophages (MDMs) were generated from normal peripheral blood mononuclear cells (PBMC). PBMCs from healthy donor were isolated by Ficoll-Hypaque gradient centrifugation. CD14 + monocytes were purified by positive selection using anti-CD14 monoclonal antibody-coated magnetic microbeads (Miltenyi Biotech, Auburn, CA) and differentiated in presence of 1 pg/ml M-CSF and 1 × 10⁶ IU/ml GM-CSF (R&D Systems) as described previously [25 (link)]. Half the volume of media was replaced every third day with fresh media containing GM-CSF and M-CSF for 7–8 days to differentiate them into MDMs.

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Reis R.S., Wagner M.C., McKenna S, & Ayyavoo V. (2024). Neuroinflammation driven by Human Immunodeficiency Virus-1 (HIV-1) directs the expression of long noncoding RNA RP11–677M14.2 resulting in dysregulation of Neurogranin in vivo and in vitro. Research Square.

Publication Preprint 2024

anti-CD14 monoclonal antibody-coated magnetic microbeads M-CSF GM-CSF

Corresponding Organization :

Other organizations : University of Pittsburgh

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Variable analysis

independent variables

M-CSF concentration (1 pg/ml)
GM-CSF concentration (1 × 10^6 IU/ml)

dependent variables

Differentiation of CD14+ monocytes into monocyte-derived macrophages (MDMs)

control variables

Peripheral blood mononuclear cells (PBMCs) from healthy donors
CD14+ monocyte purification using anti-CD14 monoclonal antibody-coated magnetic microbeads
Differentiation duration (7-8 days)
Media replacement every third day with fresh media containing GM-CSF and M-CSF

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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