Human peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by ficoll Hypaque density-gradient separation. T cells were purified by negative magnetic separation using magnetic beads containing antibodies against CD19, CD20, CD14, CD56 (Pan T-Cell Isolation Kit, Miltenyi Biotech, Auburn, CA, USA). LAN-1 tumor cells were obtained from Children’s Hospital Los Angeles. JN-DSRCT tumor cells were obtained from Fukuoka University, Fukuoka, Japan. Tap-deficient HLA-A2+ T2 cells, NK-92-MI and all other cell lines used were purchased from ATCC (Manassas, VA, USA) or developed at Memorial Sloan Kettering Cancer Center. Cells were cultured in RPMI 1640 with 2 mM L-glutamine and 10% fetal bovine serum. All cell lines have been tested and authenticated by short tandem repeat profiling using PowerPlex 10 System (Promega, Fitchburg, WI, USA) and tested for mycoplasma contamination. NK-92-MI cells and genetically CAR-modified NK-92-MI cells were propagated in Alpha Minimum Essential medium with 2 mM L-glutamine, 1.5 g/L sodium bicarbonate, 0.2 mM inositol, 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid, 12.5% horse serum and 12.5% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).
Nk 92mi
Manufactured by Thermo Fisher Scientific
Sourced in United States
The NK-92MI is a human natural killer cell line derived from a patient with non-Hodgkin's lymphoma. It is a well-characterized model system for studying natural killer cell biology and function. The NK-92MI cell line retains the key characteristics of primary natural killer cells, including the expression of natural killer cell-associated receptors and the ability to exhibit cytotoxic activity against various target cells.
Automatically generated - may contain errors
Lab products found in correlation
Nk 92mi, by American Type Culture Collection (9 mentions)
L glutamine, by Thermo Fisher Scientific (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Horse serum, by Thermo Fisher Scientific (5 mentions)
Folic acid, by Merck Group (5 mentions)
Inositol, by Merck Group (5 mentions)
Alpha minimum essential medium, by Thermo Fisher Scientific (4 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (4 mentions)
Mda mb 231, by American Type Culture Collection (3 mentions)
Mda mb 231, by Korean Cell Line Bank (3 mentions)
Sodium bicarbonate, by Merck Group (2 mentions)
Sodium bicarbonate, by Thermo Fisher Scientific (2 mentions)
2 mercaptoethanol, by Merck Group (2 mentions)
Ea hy926, by American Type Culture Collection (2 mentions)
Mcf 7, by Korean Cell Line Bank (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Mda mb 231, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Pan t cell isolation kit, by Miltenyi Biotec (1 mentions)
15 protocols using nk 92mi
1
Isolation and Cultivation of Diverse Cell Lines
2
Cultivation of NK-92 and NK-92MI Cells
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NK-92 and NK-92MI (which was virally transduced to stably express IL-2) cells were purchased from ATCC. NK-92MI cells were maintained in MEMα (Invitrogen, Carlsbad, CA, United States) supplemented with 12.5% horse serum (gibco, United States), 12.5% fetal bovine serum (gibco, United States), 0.2 mM inositol, 0.1 mM β-mercaptoethanol, 0.02 mM folic acid and penicillin/streptomycin. NK-92 was cultured based on the NK-92MI culture medium, and 100–200 U/ml recombinant IL-2 was added. Human ovarian cancer cell lines SKOV3 were maintained in DMEM basic supplemented with 10% fetal bovine serum and penicillin/streptomycin.
3
Culturing and Characterizing NK and Prostate Cancer Cells
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NK-92MI (human NK cell line) and PC3 (human prostate cancer cell line) TrampC1 (mouse prostate cancer cell line) were purchased from the American Type Culture Collection (ATCC). NK-92MI cells were cultured in Minimum Essential Medium alpha (Invitrogen) supplemented with 2 mM L-glutamine (Invitrogen), 1.5 g/L sodium bicarbonate (Sigma-Aldrich), 0.2 mM inositol (Sigma-Aldrich), 0.1 mM 2-mercaptoethanol (Invitrogen), 0.02 mM folic acid (Sigma-Aldrich), 12.5% fetal bovine serum, and 1% penicillin/streptomycin. PC3 cells were grown in standard RPMI medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (Invitrogen). TrampC1 cells were cultured in Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 0.005 mg/ml bovine insulin and 10 nM dehydroisoandrosterone, 90%; fetal bovine serum, 5%; Nu-Serum IV, 5%. Mouse primary NK cells were purified from spleen and characterized by flow cytometry analysis.
4
Cell Culture Conditions for Diverse Cell Lines
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The cell lines C4‐2, PC‐3, LNCaP, SKVO3, NK92MI and 293T were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). The NK92MI cell line is an interleukin‐2 (IL‐2)‐independent NK‐cell line derived from the NK92 cell line via transfection with retrovirus MFG‐hIL‐2 vector. The cancer cell lines (C4‐2, PC‐3, LNCaP, and SKVO3) were grown in RPMI 1640 medium (Gibco, Grand Island, NY, USA) supplemented with 1% penicillin/streptomycin (P/P; HyClone, Logan, UT, USA), and 10% fetal bovine serum (FBS; Bioind, Kibbutz Beit Haemek, Israel) at 37°C in a humidified atmosphere with 5% CO2. 293T cells were maintained in dulbecco's modified eagle medium (DMEM, HyClone) supplemented with 10% FBS and 1% P/P. NK92MI cells were cultured with minimum essential medium α (MEMα) (Invitrogen, Carlsbad, CA, USA) supplemented with 12.5% horse serum (Gibco), 12.5% FBS, 0.1 mmol/L β‐mercaptoethanol, 0.2 mmol/L inositol, and 1% P/P.
5
Culturing Human NK and Breast Cancer Cells
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NK-92MI and MDA-MB231 cells were purchased from the American Type Culture
Collection (ATCC, Manassas, VA, USA). The human NK cell line NK-92MI was
cultured in Minimum Essential Medium alpha (MEM-α,
Invitrogen, Carlsbad, CA, USA) supplemented with 2 mM L-glutamine (Invitrogen),
1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM
myo-inositol (Sigma-Aldrich), 0.1 mM 2-mercaptoethanol (Invitrogen), 0.02 mM
folic acid (Sigma-Aldrich), 12.5% fetal bovine serum (Invitrogen) and
1% penicillin/streptomycin (Invitrogen). XRP44X was purchased from
Sigma-Aldrich and dissolved in dimethyl sulfoxide (DMSO, Invitrogen).
Collection (ATCC, Manassas, VA, USA). The human NK cell line NK-92MI was
cultured in Minimum Essential Medium alpha (MEM-α,
Invitrogen, Carlsbad, CA, USA) supplemented with 2 mM L-glutamine (Invitrogen),
1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM
myo-inositol (Sigma-Aldrich), 0.1 mM 2-mercaptoethanol (Invitrogen), 0.02 mM
folic acid (Sigma-Aldrich), 12.5% fetal bovine serum (Invitrogen) and
1% penicillin/streptomycin (Invitrogen). XRP44X was purchased from
Sigma-Aldrich and dissolved in dimethyl sulfoxide (DMSO, Invitrogen).
6
Cultivation of U266 and NK-92MI Cell Lines
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The human MM cell line U266 [34 (link)] was obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). The NK cell line NK-92MI (ATCC) was cultured in MEMα medium without nucleosides (Thermo Fisher, Waltham, MA, USA) containing 12.5% fetal bovine serum, 12.5% horse serum, 0.2 mM inositol, 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid, 100 U/mL of penicillin, and 100 mg/mL of streptomycin.
7
Establishment of Radiation-Resistant Breast Cancer Cell Lines
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Human breast cancer cell lines, MDA-MB-231, MCF7, and T47D, were obtained from the Korea Cell Line Bank (Seoul, Republic of Korea). The non-malignant breast epithelial cell line, MCF10A, mouse breast cancer cell line, 4T1, human umbilical endothelial cell line, EA.hy926, and human NK cell line, NK-92MI, were obtained from American Type Culture Collection (ATCC; Manassas, VA, USA). RT-R-MDA-MB-231, RT-R-MCF7, RT-R-T47D, and RT-R-4T1 cells were generated by repetitively applying a 2 Gy dose of X-rays until a final dose of 50 Gy was achieved. All cancer cell lines were cultured in RPMI 1640 supplemented with 10% FBS, 100 IU/mL penicillin, and 10 μg/mL streptomycin, and incubated at 37 °C in a humidified atmosphere containing 5% CO2. NK-92MI cells were cultured in alpha-MEM containing 2 mM L-glutamine, 1.5 g/L sodium bicarbonate (GIBCO; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 0.2 mM inositol (Sigma-Aldrich, St. Louis, WA, USA), 0.1 mM 2-mercaptoethanol (Sigma-Aldrich, St. Louis, WA, USA), 0.02 mM folic acid (Sigma-Aldrich, St. Louis, WA, USA), 12.5% horse serum (GIBCO), and 12.5% FBS without ribonucleosides and deoxyribonucleosides at 37 °C in a humidified atmosphere containing 5% CO2.
8
Cell Culture Protocols for Cancer Research
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Vero (CCL-81), CT26WT (CRL-2638), HEK293T cells (CRL-3216) were acquired from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM, HyClone, Waltham, Massachusetts or Corning, Manassas, Virginia) with 10% FBS. JIMT-1 (ACC589) were acquired from DSMZ and cultured in DMEM with 10% FBS. 4T1.2 cells were cultured in Roswell Park Memorial Institute medium (RPMI; ATCC, Cat. # ATCC 30-2001) with 10% FBS. NK-92MI (CRL-2408) cells were cultured in RPMI with 10% FBS, 30 mM HEPES, 50 mM 2-mercaptoethanol (ThermoFisher Scientific, Cat. # 21985023).
ID8 cells were cultured in DMEM supplemented with 4% FBS and 1% ITSS (5 μg/mL insulin, 5 μg/mL transferrin, and 5 ng/mL sodium selenite; R&D Systems, MN, USA, Cat. # AR013). MC38 cells were cultured in DMEM with 10% FBS. AF2068 cells were cultured in RPMI with 10% FBS.
All media were supplemented with 1% penicillin-streptomycin solution (ThermoFisher Scientific, Cat. # 15140163). All cells were maintained at 370C in a 5% CO2 humidified incubator, routinely tested for mycoplasma contamination by Hoechst staining and PCR (Diamed, Mississauga, Ontario, Catalogue # ABMG238) and used within 3-10 passages since thaw.
ID8 cells were cultured in DMEM supplemented with 4% FBS and 1% ITSS (5 μg/mL insulin, 5 μg/mL transferrin, and 5 ng/mL sodium selenite; R&D Systems, MN, USA, Cat. # AR013). MC38 cells were cultured in DMEM with 10% FBS. AF2068 cells were cultured in RPMI with 10% FBS.
All media were supplemented with 1% penicillin-streptomycin solution (ThermoFisher Scientific, Cat. # 15140163). All cells were maintained at 370C in a 5% CO2 humidified incubator, routinely tested for mycoplasma contamination by Hoechst staining and PCR (Diamed, Mississauga, Ontario, Catalogue # ABMG238) and used within 3-10 passages since thaw.
9
Culturing and Characterizing Cancer Cell Lines
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Human cells, HT24 (bladder cancer, HTB-4, ATCC), J82 (bladder cancer, HTB-1, ATCC), BFTC909 (colorectal cancer, 60069, BCRC), H292 (lung cancer, CRL-1848, ATCC), A549 (lung cancer, CCL-185, ATCC) and NK-92MI (lymphoma, CRL-2408, ATCC) were obtained from the American Type Culture Collection (ATCC) or Bioresource Collection and Research Center (BCRC). Cells were cultured in DMEM medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% (volume/volume; v/v) fetal bovine serum (FBS), sodium bicarbonate and 1% (v/v) antibiotic-antimycotic (Thermo Fisher Scientific, Waltham, MA, USA) at 37°C in an incubator with a humidified atmosphere of 5% CO2. NK-92MI cells were cultured in alpha minimum essential medium (Thermo Fisher Scientific) supplemented with 12.5% fetal bovine serum (FBS, Sigma-Aldrich, St. Louis MO, USA), 2 mM L-glutamine (Thermo Fisher Scientific), 1.5 g/L sodium bicarbonate (Thermo Fisher Scientific), 0.2 mM inositol; (I7508, Sigma-Aldrich), 0.1 mM 2-mercaptoethanol; (M6250, Sigma-Aldrich), 0.02 mM folic acid; (F8758, Sigma-Aldrich), 12.5% horse serum. Latrunculin B (L5288, Sigma-Aldrich). Antibodies, CD161/NK1.1 was from Novus Biologicals (NB100-77528SS, Littleton, CO, USA), vimentin was from abcam (ab92547, Cambridge, UK). Phalloidin staining reagent was from Abcam (ab112125, Cambridge, UK).
10
Breast Cancer Cell Co-culture Assay
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Human breast cancer cells MCF-7 and MDA-MB-231 were obtained from the Korean Cell Line Bank (KCLB; Seoul, Korea). The human Natural Killer effector cells NK-92MI were obtained from American Type Cell Collection (ATCC; Rockville, MD, USA). Breast cancer cells were cultured at 37 °C with 5% CO2, in RPMI1640 supplemented with 2 mM l -glutamine, 10% FBS (Gibco), and 1% penicillin streptomycin (Gibco). NK-92MI cells were cultured in α-MEM supplemented with 1.5 mM sodium bicarbonate, 0.2 mM inositol, 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid, and 12.5% heat-inactivated FBS, and HS (Gibco) added in the final concentration. Since NK-92MI is an IL-2 independent cell line, fresh medium was added instead of giving complete medium change, maintaining an appropriate ratio of old and fresh medium.
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