||Human lymphoblasts from peripheral blood of a child with acute leukemia.
||This cell line is a suitable transfection host.
||Genes Expressed: CD3; Homo sapiens, expressed ,CD5; Homo sapiens, expressed ,CD7; Homo sapiens, expressed ,CD4; Homo sapiens, expressed
||RPMI-1640 medium modified to contain 2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4500 mg/L glucose, and 1500 mg/L sodium bicarbonate and fetal bovine serum to a final concentration of 10%.
||Cultures can be maintained by addition of fresh medium.
Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 10e5 viable cells/mL.
Maintain cultures at a cell concentration between 1 x 10e5 and 1 x 10e6 cells/mL.
NOTE: Do not allow the cell concentration to exceed 1 x 10e6 cells/mL.
Population Doubling Time about: 24-30 hours
||Atmosphere: air, 95%; carbon dioxide (CO2), 5%
||95% FBS + 5% DMSO (Dimethyl sulfoxide)
|Thawing Frozen Cells:
||SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that
some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris.
1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes.
4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio).
5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line).
NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
||Foley GE, et al. Continous culture of human lymphoblasts from peripheral blood of a child with acute leukemia. Cancer 18: 522-529, 1965. PubMed: 14278051
Sandstrom PA, Buttke TM. Autocrine production of extracellular catalase prevents apoptosis of the human CEM T-cell line in serum-free medium. Proc. Natl. Acad. Sci. USA 90: 4708-4712, 1993. PubMed: 8506323
Adams RA. Formal discussion: the role of transplantation in the experimental investigation of human leukemia and lymphoma. Cancer Res. 27: 2479-2482, 1967. PubMed: 4170381
Uzman BG, et al. Morphologic variations in human leukemic lymphoblasts (CCRF-CEM cells) after long-term culture and exposure to chemotherapeutic agents. A study with the electron microscope. Cancer 19: 1725-1742, 1966. PubMed: 5224274
Adams RA, et al. Leukemia: serial transplantation of human leukemic lymphoblasts in the newborn Syrian hamster. Cancer Res. 27: 772-783, 1967. PubMed: 4295047
Miranda L, et al. Isolation of the human PC6 gene encoding the putative host protease for HIV-1 gp160 processing in CD4+ T lymphocytes. Proc. Natl. Acad. Sci. USA 93: 7695-7700, 1996. PubMed: 8755538
CCRF-CEM is a T lymphoblastoid cell line derived by G.E. Foley, et al. using cells obtained in November, 1964 from peripheral blood buffy coat of a 4-year-old Caucasian female with acute lymphoblastic leukemia.
||Antonio Monteiro - Banco de Células do Rio de Janeiro