Isolation and Activation of Primary CD4+ T Cells from Rat Peripheral Blood

Primary CD4⁺ T lymphocytes were isolated from the peripheral blood mononuclear cells of SD rats using Ficoll gradient separation (GE Healthcare, Uppsala, Sweden). T lymphocytes were activated in the presence of plate-bound anti-CD3/anti-CD28 antibodies and cultivated under polarization conditions (13 (link), 14 (link)). The purity of the CD4⁺ T lymphocytes was verified using FACS analysis after their isolation and found to always remain above 90% (Supplement Figure 1). Fibroblast-like synoviocytes were isolated by enzymatic digestion of the synovial tissue from the SD rats (15 (link), 16 (link)).

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Du Y.Y., Chen Z.X., Liu M.Y., Liu Q.P., Lin C.S., Chu C.Q, & Xu Q. (2020). Leonurine Regulates Treg/Th17 Balance to Attenuate Rheumatoid Arthritis Through Inhibition of TAZ Expression. Frontiers in Immunology, 11, 556526.

Publication 2020

Ficoll gradient separation

Anti antibodies Cd4 t lymphocytes Digestion Enzymatic Fibroblast Ficoll Isolation Peripheral blood mononuclear cells Rats Supplement Synovial tissue Synoviocytes T lymphocytes

Corresponding Organization : Oregon Health & Science University

Other organizations : South China Agricultural University

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Variable analysis

independent variables

Polarization conditions (13, 14)

dependent variables

Purity of CD4+ T lymphocytes

control variables

Plate-bound anti-CD3/anti-CD28 antibodies

controls

Positive control: Activation of T lymphocytes using plate-bound anti-CD3/anti-CD28 antibodies
Negative control: Not mentioned

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