NK3.3

Species - Tissue - Features - Reference - Growth - Passage - Medium - Freeze - Source

Species: human

Tissue: bone marrow

Features: IL-2 dependent

Reference:

• Kornbluth, J., Flomenberg, N., and Dupont, B. Cell surface phenotype of a cloned line of human natural killer cells. J.Immunol. 129 (6):2831-2837, 1982.Medline
• Kornbluth J, Wilson DB: Monoclonal antibodies directed against HLA molecules affect the lytic and proliferative behavior of a cloned line of human natural killer cells. Hum Immunol 11(4), 239-247 (1984) Medline
• Kornbluth J, Spear B, Raab SS, Wilson DB: Evidence for the role of class I and class II HLA antigens in the lytic function of a cloned line of human natural killer cells. J Immunol 134(2), 728-735 (1985) Medline
• Mahle NH, Radcliff G, Sevilla CL, Kornbluth J, Callewaert DM: Kinetics of cellular cytotoxicity mediated by a cloned human natural killer cell line. Immunobiology 179(2-3), 230-243 (1989) Medline
• Zhou J, Olsen S, Moldovan J, Fu X, Sarkar FH, Moudgil VK, Callewaert DM: Glucocorticoid regulation of natural cytotoxicity: effects of cortisol on the phenotype and function of a cloned human natural killer cell line. Cell Immunol 178(2), 108-116 (1997) Medline

Mode of Growth:

• Cells grow in suspension. Feed cells every 3-4 days. Cells are maintained in T25 or T75 flasks kept upright in the incubator with 6% CO₂. The upright position favors cell-cell interaction, which appears to help them grow.

Method of Passage:

• Draw off the half to 2/3's of old medium without disturbing the cells at the bottom of the flask. Resuspend cells in the rest and count. Resuspend cells in fresh, warm medium at a concentration of 3 x 10⁵ cells per ml. Do not have a total volume exceeding 20 ml in a T25 or 50 ml in a T75 flask.

Culture Medium:

• RPMI 1640 (pH 7.2)
• 15% heat inactivated fetal bovine serum (FBS)
• 15% PHA-stimulated PBL supernatant as IL-2 source (e.g. Lymphocult-T from Biotest, No. 811010)
• 2 mM L-glutamine
• 10-25 mM HEPES
• 2 g/l Sodium Bicarbonate (NaHCO₃)
• 50-100 IU/ml Penicillin,  50-100 µg/ml Streptomycin

Freeze Medium:

Freeze cells in 90% FBS, 10% DMSO at a minimum of 10⁷ cells / ml.

Source: Dr. Jacki Kornbluth, Ph.D., Department of Pathology, Saint Louis University
School of Medicine, 1402 South Grand Blvd, Saint Louis, Missouri 63104, Tel +1 314-577-8475, kornj@slucare1.sluh.edu

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