Natural killer Cells (NK), human

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Specialized periphal blood lymphocytes that recognize and destroy foreign cells or infected host cells in a nonspecific manner.

Natural killer (NK) cells are lymphocytes of the immune system that are critical in host defense and immune regulation. Since they are part of innate immunity, they do not require sensitization for the expression of their activity. NK cells play significant roles in viral infections, autoimmunity, pregnancy, cancer, bone marrow transplantation, and more recently adaptive immunity. NK cells are most traditionally characterized by the presence of surface marker CD56.

Cell Type:
Natural Killer (NK)
Tissue Origin:
blood
Species:
human
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Research Area:
Cancer Research/Cell Biology
Immunotherapy / Hematology
Cell Characteristics:
Suspension

Recommended Media

X-VIVO

X-VIVO Serum-free Hematopoietic Cell Media provide nutritionally complete and balanced environments for a variety of cells including lymphokine activated killer (LAK) cells, peripheral blood lymphocytes (PBL), and tumor infiltrating lymphocytes (TIL).These media do not contain any exogenous growth factors, artificial stimulators of cellular proliferation, or undefined supplements. They are devoid of any protein kinase C stimulators and are suitable for the investigation of second messenger systems in the activation of human and murine lymphocytes.

There are three versions of this media:

  • X-VIVO 10  - designed to support the generation of LAK cells in a serum-free environment.
  • X-VIVO 15 - similar in composition to X-VIVO 10 but optimized for the proliferation of tumor infiltrating lymphocytes (TIL) under serum-free conditions. It also supports the proliferation of purified CD3+ cells isolated from peripheral blood and human tumors, and can be used to support the growth of human monocytes, macrophage cells and cell lines, PBL, granulocytes, and natural killer (NK) cells as well as the expansion of HUT-78 and related human lymphocytic cell lines.
  • X-VIVO 20 - optimized to support the generation of lymphokine activated killer (LAK) cells from monocyte depleted peripheral blood lymphocytes (PBL) at high density and has also been used as a growth medium for PBL and tumor infiltrating lymphocytes (TIL).

Storage = 2ºC to 8ºC

04-380Q - X-VIVO 10 with L-glutamine, gentamicin and phenol red
04-743Q - X-VIVO 10 with L-glutamine, but without gentamicin or phenol red
04-418 - X-VIVO 15  with L-glutamine, gentamicin and phenol red
04-744Q - X-VIVO 15 with L-glutamine, but without gentamicin or phenol red
04-448Q - X-VIVO 20  with L-glutamine, gentamicin and phenol red

Lymphocyte Growth Medium-3
LGM-3 is a serum-free medium for lymphocyte and dendritic cell growth

Transfection Information

Lonza Optimized Protocol
Optimization Guideline
Filter:
The table below shows data for the cell type and Nucleofector™ Platform selected. Those data are either based on Lonza Optimized Protocols or on results shared from customers who performed an optimization based on our guidelines. In case no data are shown for the selected Nucleofector™ Platform, please take a look at our optimization strategy to get further guidance on how to easily determine optimal Nucleofection conditions yourself.
Protocol Kit Program Cells Efficiency Viable Cells Substrate Format Platform
NK cell, human U-001 2e6 16% ±4 50-60% Plasmid (general) 2 µg 100 µl I/II/2b
P3 EK-100 5e5 30-32% 76% Plasmid (general) 400 ng 20 µl 4D X-Unit
P3 FA-100 5e5 53-54% 44-45% Plasmid (general) 400 ng 20 µl 4D X-Unit
P3 EN-138 3e6 35.5% 77.5% Plasmid (general) 20 µl 4D X-Unit

Citations (24)

Engineering of potent CAR NK cells using non-viral Sleeping Beauty transposition from minimalistic DNA vectors
Categories:
Primary Cells and Media, Transfection 
Authors:
Bexte T, Botezatu L, Miskey C, Gierschek F, Moter A, Wendel P, Reindl LM, Campe J, Villena-Ossa JF, Gebel V, Stein K, Cathomen T, Cremer A, Wels WS, Hudecek M, Ivics Z, Ullrich E. 
In:
Mol Ther (2024) S1525-0016: 00320-4 
Integration of ?-deficient CARs into the CD3-zeta gene conveys potent cytotoxicity in T and NK cells
Categories:
Transfection, Cell Lines and Primary Cancer Cells 
Authors:
Kath J, Franke C, Drosdek V, Du W, Glaser V, Fuster-Garcia C, Stein M, Zittel T, Schulenberg S, Porter CE, Andersch L, Künkele A, Alcaniz J, Hoffmann J, Abken H, Abou-El-Enein M, Pruß A, Suzuki M, Cathomen T, Stripecke R, Volk HD, Reinke P, Schmueck-Henneresse M, Wagner DL 
In:
Blood (2024) 143(25): :2599-2611 
Protocol for genomic editing in human resting primary NK cells and NK-92 cells via CRISPR-Cas9 ribonucleoproteins
Categories:
Transfection 
Authors:
Verhezen T, Lau HW, Van Audenaerde J, Wouters A, Smits E, De Waele J 
In:
STAR Protoc (2024) 5(3): 103123 
CRISPR/Cas9 editing of NKG2A improves the efficacy of primary CD33-directed chimeric antigen receptor natural killer cells
Categories:
Transfection 
Authors:
Bexte T, Albinger N, Al Ajami A, Wendel P, Buchinger L, Gessner A, Alzubi J, Särchen V, Vogler M, Rasheed HM, Jung BA, Wolf S, Bhayadia R, Oellerich T, Klusmann JH, Penack O, Möker N, Cathomen T, Rieger MA, Imkeller K, Ullrich E 
In:
Nat Commun. (2024) 15(1): 8439 
Peptide-mediated delivery of CRISPR enzymes for the efficient editing of primary human lymphocytes
Categories:
Primary Cells and Media, Serum-free and Speciality Media, Transfection, Cell Lines and Primary Cancer Cells 
Authors:
Foss DV, Muldoon JJ, Nguyen DN, Carr D, Sahu SU, Hunsinger JM, Wyman SK, Krishnappa N, Mendonsa R, Schanzer EV, Shy BR, Vykunta VS, Allain V, Li Z, Marson A, Eyquem J, Wilson RC 
In:
Nat Biomed Eng (2023) 5: 647-660 
The NK cell checkpoint NKG2A maintains expansion capacity of human NK cells
Categories:
Transfection 
Authors:
Kaulfuss M, Mietz J, Fabri A, Vom Berg J, Münz C, Chijioke O 
In:
Biosci Rep (2023) 13(1): 10555 
CRISPR-Cas9 based gene editing of the immune checkpoint NKG2A enhances NK cell mediated cytotoxicity against multiple myeloma
Categories:
Transfection 
Authors:
Bexte T, Alzubi J, Reindl LM, Wendel P, Schubert R, Salzmann-Manrique E, von Metzler I, Cathomen T, Ullrich E 
In:
Oncol Rep (2022) 31;11(1): 2081415 
Optimization and validation of CAR transduction into human primary NK cells using CRISPR and AAV
Categories:
Primary Cells and Media, Transfection 
Authors:
Naeimi Kararoudi M, Likhite S, Elmas E, Yamamoto K, Schwartz M, Sorathia K, de Souza Fernandes Pereira M, Sezgin Y, Devine RD, Lyberger JM, Behbehani GK, Chakravarti N, Moriarity BS, Meyer K, Lee DA 
In:
Cell Rep Med (2022) 2(6): 100236 
CRISPR/Cas9-Based Gene Engineering of Human Natural Killer Cells: Protocols for Knockout and Readouts to Evaluate Their Efficacy
Categories:
Serum-free and Speciality Media, Transfection 
Authors:
Mélanie Lambert , Caroline Leijonhufvud , Filip Segerberg , J Joseph Melenhorst , Mattias Carlsten  
In:
Methods Mol Biol (2020) 2121: 213-219 
Polymer-stabilized Cas9 nanoparticles and modified repair templates increase genome editing efficiency.
Categories:
Primary Cells and Media, Serum-free and Speciality Media, Transfection 
Authors:
Nguyen DN, Roth TL, Li PJ, Chen PA, Apathy R, Mamedov MR, Vo LT, Tobin VR, Goodman D, Shifrut E, Bluestone JA, Puck JM, Szoka FC, Marson A. 
In:
Nat Biotechnol (2019) 1: 1 
NK cells switch from granzyme B to death receptor-mediated cytotoxicity during serial killing.
Categories:
Transfection 
Authors:
Prager I, Liesche C, van Ooijen H, Urlaub D, Verron Q, Sandström N, Fasbender F, Claus M, Eils R, Beaudouin J, Önfelt B, Watzl C. 
In:
J Exp Med (2019) 216(9): 2113-2127 
Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins.
Categories:
Primary Cells and Media, Transfection 
Authors:
Naeimi Kararoudi, M., Dolatshad, H., Trikha, P., Hussain, S. R., Elmas, E., Foltz, J. A., Moseman, J. E., Thakkar, A., Nakkula, R. J., Lamb, M., Chakravarti, N., McLaughlin, K. J., Lee, D. A. 
In:
Journal of Visualized Experiments, JoVE (2018) 136: e58237 
Enhanced NK cell adoptive antitumor effects against breast cancer in vitro via blockade of the transforming growth factor-ß signaling pathway.
Categories:
Transfection 
Authors:
Zhao Y1, Hu J2, Li R1, Song J1, Kang Y1, Liu S1, Zhang D1. 
In:
OncoTarget (2015) 8: 1553-9 
Targeting CD20+ Aggressive B-cell Non-Hodgkin Lymphoma by Anti-CD20 CAR mRNA-Modified Expanded Natural Killer Cells In Vitro and in NSG Mice.
Categories:
Transfection 
Authors:
Chu Y, Hochberg J, Yahr A, Ayello J, van de Ven C, Barth M, Czuczman M, Cairo MS. 
In:
Cancer Immunol Immunother (2015) 3(4): 333-44 
Age-associated changes in the differentiation potentials of human circulating hematopoietic progenitors to T- or NK-lineage cells
Categories:
Primary Cells and Media 
Authors:
Kyoizumi S, Kubo Y, Kajimura J, Yoshida K, Imai K, Hayashi T, Nakachi K, Young LF, Moore MA, van den Brink MR, Kusunoki Y. 
In:
J Immunol (2013) 190(12): 6164-72 
Human leukocyte antigen I knockdown human embryonic stem cells induce host ignorance and achieve prolonged xenogeneic survival
Categories:
Primary Cells and Media 
Authors:
Deuse T, Seifert M, Phillips N, Fire A, Tyan D, Kay M, Tsao PS, Hua X, Velden J, Eiermann T, Volk HD, Reichenspurner H, Robbins RC, Schrepfer S. 
In:
Circulation (2011) 124(11): ePub 
NKG2D ligand expression in AML increases in response to HDAC inhibitor valproic acid and contributes to allorecognition by NK-cell lines with single KIR-HLA class I specificities.
Categories:
Classical Media and Reagents, Serum-free and Speciality Media 
Authors:
Diermayr S, Himmelreich H, Durovic B, Mathys-Schneeberger A, Siegler U, Langenkamp U, Hofsteenge J, Gratwohl A, Tichelli A, Paluszewska M, Wiktor-Jedrzejczak W, Kalberer CP, Wodnar-Filipowicz A. 
In:
Blood (2008) 111(3): 1428-36 
Regulation of human DAP10 gene expression in NK and T cells by Ap-1 transcription factors
Categories:
Transfection 
Authors:
Marusina AI, Burgess SJ, Pathmanathan I, Borrego F, Coligan JE 
In:
J Immunol (2008) 180(1): 409-17 
Differential regulation of human NK cell-mediated cytotoxicity by the tyrosine kinase itk
Categories:
Transfection 
Authors:
Khurana D, Arneson LN, Schoon RA, Dick CJ, Leibson PJ 
In:
J Immunol (2007) 178(6): 3575-82 
Nucleofection of non-B cells with mini-Epstein-Barr virus DNA
Categories:
Transfection 
Authors:
Radons J, Gross C, Stangl S and Multhoff G 
In:
J Immunol Methods (2005) 303(1-2): 135-141 
Three Structurally and Functionally Divergent Kinds of Promoters Regulate Expression of Clonally Distributed Killer Cell Ig-Like Receptors (KIR), of KIR2DL4, and of KIR3DL3
Categories:
Transfection 
Authors:
Trompeter HI, Gomez-Lozano N, Santourlidis S, Eisermann B, Wernet P, Vilches C and Uhrberg M 
In:
J Immunol (2005) 174(7): 4135-4143 
Activated human NK and CD8+ T cells express both TNF-related apoptosis-inducing ligand (TRAIL) and TRAIL receptors but are resistant to TRAIL-mediated cytotoxicity
Categories:
Transfection 
Authors:
Mirandola P, Ponti C, Gobbi G, Sponzilli I, Vaccarezza M, Cocco L, Zauli G, Secchiero P, Manzoli FA and Vitale M 
In:
Blood (2004) 104(8): 2418-2424 
Dynamic redistribution of the activating 2B4/SAP complex at the cytotoxic NK cell immune synapse
Categories:
Transfection 
Authors:
Roda-Navarro P, Mittelbrunn M, Ortega M, Howie D, Terhorst C, Sanchez-Madrid F and Fernandez-Ruiz E 
In:
J Immunol (2004) 173(6): 3640-3646 
Rapid and highly efficient gene transfer into natural killer cells by nucleofection
Categories:
Transfection 
Authors:
Trompeter HI, Weinhold S, Thiel C, Wernet P and Uhrberg M 
In:
J Immunol Methods (2003) 274: 245-256 
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