Isolation and Activation of Human CD4+ T Cells

Human CD4⁺ T cells were isolated from buffy coats of healthy donors (supplied by Blodbanken, Oslo, Norway) using the Dynabeads CD4 Positive Isolation kit (Life Technologies AS, Oslo Norway) as previously described [22 (link)]. Buffy coats were diluted 1 : 2 with RPMI 1640 (Sigma–Aldrich, St. Louis, MO, U.S.A.) and 1 ml EDTA and rotated for 15 min at 4°C. CD4⁺ T cells were then isolated according to the company protocol. Cells were maintained at 5 × 10⁶ cells/ml unless otherwise stated. CD4⁺ T-cell activation was initiated by adding pre-washed Human T-Activator CD3/CD28 Dynabeads at a 1 : 4 bead/cells ratio (Life Technologies).

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Wik J.A., Chowdhury A., Kolan S., Bastani N.E., Li G., Alam K., Grimolizzi F, & Skålhegg B.S. (2022). Endogenous glutamine is rate-limiting for anti-CD3 and anti-CD28 induced CD4+ T-cell proliferation and glycolytic activity under hypoxia and normoxia. Biochemical Journal, 479(11), 1221-1235.

Publication 2022

Dynabeads CD4 Positive Isolation kit RPMI 1640 Human T-Activator CD3/CD28 Dynabeads

Cd4 t cell Cells Donors Edta Human Isolation

Corresponding Organization :

Other organizations : University of Oslo

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Variable analysis

independent variables

Activation of CD4+ T cells using Human T-Activator CD3/CD28 Dynabeads

dependent variables

Not explicitly mentioned

control variables

Maintenance of CD4+ T cells at 5 × 10^6 cells/ml
Use of buffy coats from healthy donors

controls

Positive control: Activation of CD4+ T cells using Human T-Activator CD3/CD28 Dynabeads
Negative control: Not explicitly mentioned

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