Delectinized Interleukin-2

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Catalog Number: 
03-020-050
Status: 
Current product--usually ships within 2 business days
Size: 
50 mL
Use: 
For research use only. Not for use in diagnostic procedures.
Storage Temperature: 
-70˚C

NOTE: Not for human or diagnostic use.

Procedure for Use of Human T-Cell Growth Factor (TCGF)

  1. Prepare isolated human T-lymphocytes from blood using a cell separation medium such as Histopaque or lymphocytic separation medium (LSM). Wash T-cells 3 times in RPMI-1640 fetal bovine serum 10%.
  2. Expose T-cells at a concentration of 1 × 106 cells per mL to a mitogen such as phytohemagglutinin (PHA) to activate cells for TCGF-dependent growth in vitro. Incubate activated cells 5 days at 37°C in 5% CO2.
  3. Wash cells 3 times in RPMI-1640 fetal bovine serum 10%. Resuspend cells to a concentration of 5 × 105 cells/mL in RPMI-1640 supplemented with 10 to 20% fetal bovine serum and 10% volume of TCGF, Delectinized.
  4. To split cultures, pellet cells and resuspend back to 4 to 6 × 105 cells/mL every 3 to 5 days using fresh RPMI-1640 fetal bovine serum 10 to 20% plus a 10% volume of TCGF, Delectinized.
  5. To establish long-term cultures of T-lymphocytes, repeat step 4 as necessary.

Production of TCGF: Human T-cell growth factor (TCGF) is prepared from pooled human blood leukocytes stimulated with purified phytohemagglutinin (PHA). Following induction with PHA, the leukocytes are incubated for 40 to 72 hours in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 5 mg/mL human serum albumin and 10 µg/mL Gentamicin and the crude TCGF fluids harvested.

Delectinization: TCGF fluids are clarified by centrifugation and delectinized by immunoaffinity adsorption. The final product is assayed by a peripheral blood lymphocyte mitogenic assay and found to be free of detectable levels of PHA.

Quality Control: Each lot of TCGF is screened and found to be free of bacteria, yeasts, molds and mycoplasma by accepted laboratory methods. The product is lectin-free and capable of stimulating long-term growth of adult human T-lymphocytes when assayed by long-term proliferation assays in vitro.

TCGF Assay: Human T-Cell Growth Factor, Delectinized, when used at 10% by volume in RPMI-1640 with 10% fetal bovine serum, will support the exponential growth of TCGF dependent human T-lymphocytes from 3.0 × 105 to 1.8 × 106 cells per mL in 72 hours.

Specific Activity: 50 half-maximal units/mL.