Monocyte-derived dendritic cell protocol

CD14+ cells within PBMCs were isolated using anti-CD14 magnetic microbeads and LS column separation (Miltenyi Biotech) per the manufacturer’s instructions. These cells were then cultured in complete RPMI media (10% FBS and 2 mM L-Glutamax) for 7 days in the presence of GM-CSF (20 ng/mL) and IL-6 (20 ng/mL). GM-CSF was added on days 1, 3, and 5, whereas IL-6 was added only on day 533 34 (link) (online supplemental figure 1). The cells were then harvested on day 7, and flow cytometry was performed to assess the phenotype (CD33⁺CD11b⁺HLA-DR^lowCD14⁺CD15^-).

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Nalawade S.A., Shafer P., Bajgain P., McKenna M.K., Ali A., Kelly L., Joubert J., Gottschalk S., Watanabe N., Leen A., Parihar R., Vera Valdes J.F, & Hoyos V. (2021). Selectively targeting myeloid-derived suppressor cells through TRAIL receptor 2 to enhance the efficacy of CAR T cell therapy for treatment of breast cancer. Journal for Immunotherapy of Cancer, 9(11), e003237.

Publication 2021

anti-CD14 magnetic microbeads LS column separation

Cd11b Cells Cultured media Flow cytometry Gm csf Microbeads Phenotype

Corresponding Organization : Baylor College of Medicine

Other organizations : Frederick National Laboratory for Cancer Research, National Cancer Institute, St. Jude Children's Research Hospital

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Variable analysis

independent variables

GM-CSF (20 ng/mL)
IL-6 (20 ng/mL)

dependent variables

Phenotype of cells (CD33+CD11b+HLA-DRlow CD14+CD15-)

control variables

Culture media (complete RPMI media with 10% FBS and 2 mM L-Glutamax)
Cell isolation (using anti-CD14 magnetic microbeads and LS column separation)
Cell culture duration (7 days)

controls

No explicit mention of positive or negative controls.

Annotations

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