NK-92

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

Species: human

Tissue: bone marrow

Features: IL-2 dependent

Reference:

• Brooks, A.G., Posch, P.E., Scorzelli, C.J., Borrego, F., and Coligan, J.E. NKG2A complexed with CD94 defines a novel inhibitory natural killer cell receptor. J.Exp.Med. 185 (4):795-800, 1997. Medline
• Gong, J.H., Maki, G., and Klingemann, H.G. Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia 8 (4):652-658, 1994. Medline
• Klingemann, H.G., Wong, E., and Maki, G. A cytotoxic NK-cell line (NK-92) for ex vivo purging of leukemia from blood. Biol.Blood Marrow Transplant. 2 (2):68-75, 1996. Medline
• Selvakumar, A., Steffens, U., and Dupont, B. NK cell receptor gene of the KIR family with two IG domains but highest homology to KIR receptors with three IG domains. Tissue Antigens. 48 (4 Pt 1):285-294, 1996. Medline

Mode of Growth: Cells grow in suspension. High background of cellular debris which increases with cell density. Cells grow in aggregates of 3-10 cells.

Method of Passage: Half-media change every 3-4 days. Simply let the flasks stand upright for 2 minutes. Proliferating cell aggregates sink to the bottom and the old media including most of the debris can be aspirated. Resuspend cells in the rest, count and ajust to 0.1 to 0.5 x 10⁶ cells /ml with fresh media.

Culture Medium:

1. Myelocult H5100 (#HCC-5100, Cell Systems, St.Katharinen, Tel.: +49 (2644) 9512-0)
   • 12.5% horse serum, 12.5% FBS, 0.2 mM i-inositol, 20 mM folic acid, 10^-4 M 2-mercaptoethanol, 2 mM L-glutamine, alpha MEM
   • add 100 IU/ml IL-2 (10 - 1000 IU/ml)
2. RNK-medium
3. IMDM (Iscove's modified Dulbecco's MEM)
   • 10% FBS
   • add 100 IU/ml IL-2 (10 - 1000 IU/ml)
4. RPMI 1640
   • 10% FBS
   • add 100 IU/ml IL-2 (10 - 1000 IU/ml)

Freeze Medium: culture medium + 10% DMSO, freeze 10⁷ cells / ml

Source:

• Dr. Hans G. Klingemann, M.D.
  Bone Marrow Transplant Center
  Rush Presbyterian Saint Lukes Med. Ctr.
  1653 W. Congress Pkwy.
  Chicago, IL 60612
  Tel.: +1 (312) 942-6173
  Fax.: +1 (312) 733-1590
  E-mail: hklingem@rush.edu
• ImmuneMedicine Inc.
  Tel.: +1 (312) 243-1754
• Dr. Torsten Tonn
  Institut für Transfusionsmedizin und Immunhämatologie
  BSD Hessen
  Sandhofstr. 1
  60528 Frankfurt am Main
  Tel.: +49 (69) 678-2228
  Fax.: +49 (69) 678-2259
  E-mail: ttonn@bsdhessen.de

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