|Cell Line:||HuT 78|
Vulgar Name: Human
|Age/Ethinicy:||53 years adult; Caucasian|
|Applications:||This cell line is a suitable transfection host.|
|Tumor Formation:||Yes, in nude mice when injected intracranially.|
|Products:||Genes Expressed: interleukin 2, tumor necrosis factor alpha (TNF alpha), CD4; Homo sapiens. Antigen Expression: CD4; Homo sapiens Receptor Expression: interleukin 2 (IL-2), expressed|
|Culture Medium:||Iscove's Modified Dulbecco's Medium (IMDM) contains 4 mM L-glutamine, 4500 mg/L glucose, and 1500 mg/L sodium bicarbonate. Fetal bovine serum to a final concentration of 20%.|
Cultures can be maintained by addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension in fresh medium at 2 X 105 viable cells/ml. Maintain cultures at cell concentrations between 5 X 104 and 8 X 105 viable cells/mL. Maintain cell density at less than 1 X 106 cells/mL.
NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010.
Population Doubling Time about 65 hours.
Medium Renewal: Two to three times weekly
|Culture Conditions:||Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C.|
|Cryopreservation:||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).|
|References:||Gootenberg JE, et al. Human cutaneous T cell lymphoma and leukemia cell lines produce and respond to T cell growth factor. J. Exp. Med. 154: 1403-1418, 1981. PubMed: 6975346 Mann DL, et al. Origin of the HIV-susceptible human CD4+ cell line H9. AIDS Res. Hum. Retroviruses 5: 253-255, 1989. PubMed: 2567177 Gazdar AF, et al. Mitogen requirements for the in vitro propagation of cutaneous T-cell lymphomas. Blood 55: 409-417, 1980. PubMed: 6244013 Chen TR. Karyotypic derivation of H9 cell line expressing human immunodeficiency virus susceptibility. J. Natl. Cancer Inst. 84: 1922-1926, 1992. PubMed: 1460674 O'Connell MA, et al. Cellular proliferation and activation of NF kappa B are induced by autocrine production of tumor necrosis factor alpha in the human T lymphoma line HuT 78. J. Biol. Chem. 270: 7399-7404, 1995. PubMed: 7706285 Hu SX, et al. Development of an adenovirus vector with tetracycline-regulatable human tumor necrosis factor alpha gene expression. Cancer Res. 57: 3339-3343, 1997. PubMed: 9269991 Kolanus W, et al. alphaLbeta2 integrin/LFA-1 binding to ICAM-1 induced by cytohesin-1 a cytoplasmic regulatory molecule. Cell 86: 233-242, 1996. PubMed: 8706128 Bloom TJ, Beavo JA. Identification and tissue-specific expression of PDE7 phosphodiesterase splice variants. Proc. Natl. Acad. Sci. USA 93: 14188-14192, 1996. PubMed: 8943082 Hut 78 was derived from peripheral blood of a patient with Sezary syndrome. The line has the properties of a mature human T cell with helper/inducer activity. It releases IL-2, and has receptors for IL-2. The growth rate is stimulated by IL-2. TNF alpha is an autocrine growth factor for Hut 78.|