Isolated islets pooled from eight rats were cultured with interleukin (IL)-2 to promote the expansion of activated T cells. Culture medium contained RPMI 1640 with 10% FBS, 1 mmol/L Na pyruvate, nonessential amino acids, 28 μmol/L β-mercaptoethanol, and 50 units/mL recombinant human IL-2 (PeproTech, Rocky Hill, NJ). Fifty islets per well were cultured in 24-well plates (Greiner Bio-One) in 5% CO2 at 37°C. On day 7, islets and infiltrating cells were collected and passed through a 40-μm strainer to retain the islets. T cells were pelleted and resuspended in TRIzol (28 (link)).
Protocol detail
Isolate and Expand Activated T Cells
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Amino acids
Cells
Culture medium
Interleukin 2
Mercaptoethanol
Pyruvate
Rats
Recombinant human interleukin 2
T cells
Trizol
Corresponding Organization :
Other organizations : Drexel University, University of Massachusetts Chan Medical School
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Variable analysis
independent variables
- Interleukin (IL)-2
- Expansion of activated T cells
- RPMI 1640 with 10% FBS
- 1 mmol/L Na pyruvate
- Nonessential amino acids
- 28 μmol/L β-mercaptoethanol
- 50 units/mL recombinant human IL-2
- Cultured in 24-well plates in 5% CO2 at 37°C
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