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CRL-2408 NK-92MI 人恶性非霍奇金淋巴瘤患者的自然杀伤细胞
Age:
50 years
Gender:
male
Ethnicity:
Caucasian, White
Comments:
NK-92 is an interleukin-2 (IL-2) dependent Natural Killer Cell line derived from peripheral blood mononuclear cells from a 50 year old Caucasian male with rapidly progressive non-Hodgkin's lymphoma. NK-92MI is an interleukin-2 (IL-2) independent Natural Killer Cell line derived from the NK-92 (ATCC CRL-2407) cell line by transfection. The parental cells were transfected with human IL-2 cDNA in the retroviral MFG-hIL-2 vector by particle-mediated gene transfer. The transfection is stable, most likely due to integration of the vector into genomic DNA. The cell line is cytotoxic to a wide range of malignant cells; it kills both K562 cells and Daudi cells in chromium release assays. NK-92 and derivative cell line NK-92MI have the following characteristics: surface marker positive for CD2, CD7, CD11a, CD28, CD45, CD54 and CD56 bright; surface marker negative for CD1, CD3, CD4, CD5, CD8, CD10, CD14, CD16, CD19, CD20, CD23, CD34 and HLA-DR. The parental IL-2 dependent cell line is available as CRL-2407 (NK-92). NK-92MI was shown to contain, express, and synthesize the hIL-2. A culture submitted to the ATCC in September of 1998 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline. The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.
Propagation:
ATCC complete growth medium: The base medium for this cell line is Alpha Minimum Essential medium without ribonucleosides and deoxyribonucleosides but with 2 mM L-glutamine and 1.5 g/L sodium bicarbonate . To make the complete growth medium, add the following components to the base medium: 0.2 mM inositol; 0.1 mM 2-mercaptoethanol; 0.02 mM folic acid; horse serum to a final concentration of 12.5%; fetal bovine serum to a final concentration of 12.5%. Temperature: 37.0°C
Vessels with plug-seal caps must be used to prevent evaporation of the culture medium. When using vessels with plug-seal caps, gas the headspace of the vessels with 5% ± 1% CO2. Either a standard incubator or a 5.0% ± 1.0% CO2 incubator may then be used.
Subculturing:
Protocol: CRL-2408 cell aggregates must be dispersed every 2 to 3 days by pipetting up and down on the back of the flask to produce a single cell suspension. Cell counts should be performed every 2 to 3 days to determine the viable cell density.Centrifugation and replacement of culture medium may be performed if needed to achieve the appropriate cell density.Maintain cell density between 2 X 10 (5) and 1 X 10 (6) viable cells/ml.
38894: Gong JH, et al. Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia 8: 652-658, 1994. PubMed:
38969: Tam YK, et al. Characterization of genetically altered, interleukin 2-independent natural killer cell lines suitable for adoptive cellular immunotherapy. Hum. Gene Ther. 10:
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