Rapid Expansion of Primary Immune Cells

Tissues were dissected into fragments of approximately 2 mm³. Each fragment was plated individually in a single well of a 24-well plate and stimulated with 6000 IU/mL rhIL-2 (Roche, Basel, Switzerland) for 3 weeks (Pre-REP). A rapid expansion protocol (REP) [15 (link),19 (link)] was performed by stimulating PILs with PHA 1 µg/mL (Sigma, St. Loius, MI, USA), 3000 IU/mL rhIL2, and feeders. PILs culture media was RPMI supplemented with 5% penicillin-streptomycin (Gibco, Billings, MT, USA), 25 mM HEPES, 1% L-glutamine (Gibco), 1% nonessential amino acids (Gibco), 1% Na pyruvate (Gibco), 0.1% 2β-mercaptoethanol (Gibco), and 8% heat-inactivated human serum.

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Ahmed R., Lozano L.E., Anastasio A., Lofek S., Mastelic-Gavillet B., Navarro Rodrigo B., Nguyen S., Dartiguenave F., Rodrigues-Dias S.C., Cesson V., Valério M., Roth B., Kandalaft L.E., Redchenko I., Hill A.V., Harari A., Romero P., Derré L, & Viganó S. (2023). Phenotype and Reactivity of Lymphocytes Expanded from Benign Prostate Hyperplasic Tissues and Prostate Cancer. Cancers, 15(12), 3114.

Publication 2023

rhIL-2 PHA penicillin-streptomycin L-glutamine nonessential amino acids Na pyruvate 2β-mercaptoethanol

Amino acids Culture media Glutamine Hepes Human Mercaptoethanol Penicillin Pyruvate Serum Streptomycin Tissues

Corresponding Organization : University of Lausanne

Other organizations : Ludwig Cancer Research, Jenner Institute, University of Oxford

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Variable analysis

independent variables

Tissue fragment size (approximately 2 mm^3)
Stimulation with 6000 IU/mL rhIL-2 for 3 weeks (Pre-REP)
Stimulation with PHA 1 μg/mL, 3000 IU/mL rhIL2, and feeders (REP)

dependent variables

Cell growth and proliferation during the Pre-REP and REP phases

control variables

Culture media (RPMI supplemented with 5% penicillin-streptomycin, 25 mM HEPES, 1% L-glutamine, 1% nonessential amino acids, 1% Na pyruvate, 0.1% 2β-mercaptoethanol, and 8% heat-inactivated human serum)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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