Isolation and Culture of CD4+ Cells

Cells were isolated and cultured as described previously.^{47 (link)} Assessment of CD4⁺ cell purity is demonstrated in Figure I in the online-only Data Supplement. Splenocytes were cultured in a 24-well plate (2×10⁶ cells/2 mL per well). MACSibead mouse T-cell, CD3 and CD28 antibody coated, expansion beads (Miltenyi 130-093-627) were added to medium as described.^{47 (link)} TA-65 activator (TA65) is a telomerase activator purified from Astragalus membranaceous^{52 (link)} and provided by TA-Science Inc (New York, NY). BIBR 1532 (Tocris Bioscience), a telomerase inhibitor,^{53 (link)} was dissolved in dimethyl sulfoxide and used as the indicated concentration.

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Richardson G.D., Sage A., Bennaceur K., Al Zhrany N., Coelho-Lima J., Dookun E., Draganova L., Saretzki G., Breault D.T., Mallat Z, & Spyridopoulos I. (2018). Telomerase Mediates Lymphocyte Proliferation but Not the Atherosclerosis-Suppressive Potential of Regulatory T-Cells. Arteriosclerosis, Thrombosis, and Vascular Biology, 38(6), 1283-1296.

Publication 2018

BIBR 1532

Antibody Astragalus Bibr 1532 Cd4 cell Cells Dimethyl sulfoxide Mouse Supplement T cell Telomerase

Corresponding Organization :

Other organizations : University of Cambridge, Centre for Life, Newcastle Hospitals - Campus for Ageing and Vitality, Newcastle University, Harvard Stem Cell Institute, Boston Children's Hospital, Harvard University, Sorbonne Paris Cité, Université Paris Cité, Paris Cardiovascular Research Center, Inserm, Délégation Paris 5

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Variable analysis

independent variables

TA-65 activator (TA65)
BIBR 1532 (Tocris Bioscience)

dependent variables

Assessment of CD4+ cell purity

control variables

Cells were isolated and cultured as described previously.
Splenocytes were cultured in a 24-well plate (2×10^6 cells/2 mL per well).
MACSibead mouse T-cell, CD3 and CD28 antibody coated, expansion beads (Miltenyi 130-093-627) were added to medium as described.

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