Isolation and Characterization of Resting CD4+ T Cells

Fifty milliliters of blood samples were taken from all study patients. Briefly, fresh PBMCs were obtained and rCD4+ T cells were purified by negative selection with EasySep™ Human Resting CD4+ T-Cell Isolation Kit (Stemcell Technologies, Vancouver, Canada), as described previously by our group [9 (link)]. The purity of CD4+CD25-CD69-HLA/DR-T cells was assessed by flow cytometry as >99%. Total RNA was extracted using AllPrep DNA/RNA Mini Kit (Qiagen, Hilden, Germany) from both rCD4 T+ cells and rCD4 T- PBMCs, following manufacturer’s instructions. RNA was quantified with Nanodrop 1000 (Thermo Fisher Scientific, Waltham, MA, USA).

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Martínez-Román P., López-Huertas M.R., Crespo-Bermejo C., Arca-Lafuente S., Cortegano I., Valle-Millares D., Gaspar M.L., Martín-Carbonero L., Domínguez-Domínguez L., Ryan P., de los Santos I., de la Fuente-Moral S., Fernández-Rodríguez A., Coiras M, & Briz V. (2020). Hepatitis C Virus Influences HIV-1 Viral Splicing in Coinfected Patients. Journal of Clinical Medicine, 9(7), 2091.

Publication 2020

EasySep™ Human Resting CD4+ T-Cell Isolation Kit AllPrep DNA/RNA Mini Kit Nanodrop 1000

Blood Cd25 Cd4 t cells Flow cytometry Hla dr Human Isolation Patients Rcd4 Stemcell T cells

Corresponding Organization : Hospital Universitario Ramón y Cajal

Other organizations : Centro Nacional de Microbiologia, Hospital La Paz Institute for Health Research, Hospital Universitario Infanta Leonor, Hospital Universitario de La Princesa, Hospital Universitario Puerta de Hierro Majadahonda

Top 5 similar protocols

Variable analysis

independent variables

Negative selection of rCD4+ T cells using EasySep™ Human Resting CD4+ T-Cell Isolation Kit

dependent variables

Purity of CD4+CD25-CD69-HLA/DR-T cells assessed by flow cytometry
Total RNA extracted from rCD4 T+ cells and rCD4 T- PBMCs
RNA quantification using Nanodrop 1000

control variables

Fifty milliliters of blood samples taken from all study patients
Fresh PBMCs obtained

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