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Kelly cells

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KELLY cells are a type of cell line used in laboratory research. They are derived from human neuroblastoma cells and are commonly used as a model system for studying various biological processes. The KELLY cell line maintains a stable phenotype and can be cultured in vitro.

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Lab products found in correlation

Sk n as, by American Type Culture Collection (4 mentions) Be 2 c, by American Type Culture Collection (3 mentions) Sk n as cells, by American Type Culture Collection (2 mentions) Sk n dz, by American Type Culture Collection (2 mentions) Sh sy5y, by American Type Culture Collection (2 mentions) Imr 90, by American Type Culture Collection (2 mentions) Anti mycn antibody, by Santa Cruz Biotechnology (2 mentions) α flag, by Thermo Fisher Scientific (2 mentions) V5 ip, by Thermo Fisher Scientific (2 mentions) Kelly cells, by Santa Cruz Biotechnology (2 mentions) Anti eya1 antibody, by ProSci (2 mentions) β actin, by Merck Group (2 mentions) Jetprime, by Polyplus Transfection (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Calf serum, by Thermo Fisher Scientific (1 mentions) Lookout mycoplasma detection kit, by Merck Group (1 mentions) Nih 3t3, by Merck Group (1 mentions) Hek293t, by Thermo Fisher Scientific (1 mentions)

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KELLY neuroblastoma cells (4 citations)

14 protocols using kelly cells

1

Culturing Human Neuroblastoma Cell Lines

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Human neuroblastoma MYCN amplified KELLY cells (Sigma-Aldrich, St. Louis, MO, USA) were maintained in culture using RPMI 1640 medium with GlutaMAX (Gibco, Grand Island, NY), 10% fetal bovine serum (FBS, Gibco, Grand Island, NY), 100 IU/mL penicillin (Gibco, Grand Island, NY) and 100 μg/mL streptomycin (Gibco, Grand Island, NY ). Human neuroblastoma non-MYCN amplified SKNAS cells (American Type Culture Collection, Manassas, VA, USA) were maintained in culture using Dulbecco’s modified Eagle’s medium (DMEM with GlutaMAX, HyClone, Logan, UT, USA), 10% FBS, 100 IU/mL penicillin, 100 μg/mL streptomycin, and 0.1 mM non-essential amino acids.
Taylor J.S., Yavuz B., Zeki J., Wood L., Ikegaki N., Coburn J., Harrington K., Shimada H., Kaplan D.L, & Chiu B. (2020). Enhancing sustained release local therapy: single versus dual chemotherapy for the treatment of neuroblastoma. Surgery, 167(6), 969-977.
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2

Cell Line Handling and Authentication

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NIH3T3, HEK293T, BE2C, and SHEP were purchased from the American Type Culture Collection. Kelly cells were purchased from Sigma-Aldrich. NB-19, SK-N-FI, and CHLA-90 cells were a gift from Y. DeClerck (Children's Hospital Los Angeles, Los Angeles, USA). NB-SD, NMB, NB69, and SK-N-SH cells were obtained from the Children’s Hospital of Philadelphia (CHOP) cell line bank. Tet21N cells were a gift from M. Schwab (German Cancer Research Center, Heidelberg, Germany). NIH3T3 cells stably expressing the empty vector Murine Stem Cell Virus (MSCV), oncogenic EN (ETV6-NTRK3), or mutant KRasV12 were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Sigma-Aldrich) with 10% bovine serum (calf serum, Gibco). All cell lines have been authenticated by short tandem repeat (STR) profiling using the AmpFLST Identifiler PCR Amplification Kit (Applied Biosystems) and were tested for mycoplasma on a regular basis using the LookOut Mycoplasma Detection Kit (Sigma-Aldrich). HEK293T cells were cultured in DMEM with 10% fetal bovine serum (FBS; Gibco). All other cell lines were cultured in RPMI 1640 supplemented with 10% FBS. All media were supplemented with 1% antibiotic-antimycotic (Gibco), and all cells were cultured at 37°C with 5% CO2.
Zhang H.F., Delaidelli A., Javed S., Turgu B., Morrison T., Hughes C.S., Yang X., Pachva M., Lizardo M.M., Singh G., Hoffmann J., Huang Y.Z., Patel K., Shraim R., Kung S.H., Morin G.B., Aparicio S., Martinez D., Maris J.M., Bosse K.R., Williams K.C, & Sorensen P.H. (2023). A MYCN-independent mechanism mediating secretome reprogramming and metastasis in MYCN-amplified neuroblastoma. Science Advances, 9(34), eadg6693.
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3

Cell Line Culture and Characterization

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BE(2)C (ATCC #CRL-2268), PA-1 (ATCC #CRL-1572), IMR90 (ATCC #CRL-186), SK-N-AS (ATCC #CRL-2137), SH-SY5Y (ATCC #CRL-2266), 293T(ATCC #11268), SK-N-DZ (ATCC #CRL-2149) and Kelly cells (Sigma #92110411-1VL) were maintained in 1:1 DMEM/F12:MEM media with 10% inactivated fetal calf serum, 1ug/ml penicillin, and 1U/ml streptomycin. All cell lines were purchased for the purposes of this study, are not among commonly misidentified cell lines (per ICLAC), and tested negative for mycoplasma contamination.
Powers J.T., Tsanov K.M., Pearson D.S., Roels F., Spina C.S., Ebright R., Seligson M., de Soysa Y., Cahan P., Theiβen J., Tu H.C., Han A., Kurek K.C., LaPier G.S., Osborne J.K., Ross S.J., Cesana M., Collins J.J., Berthold F, & Daley G.Q. (2016). Multiple mechanisms disrupt the let-7 microRNA family in neuroblastoma. Nature, 535(7611), 246-251.
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4

Culturing Human Neuroblastoma KELLY Cells

Cited 1 time
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Human neuroblastoma KELLY cells (Sigma-Aldrich, St Louis, MO) were maintained in RPMI 1640 (HyClone, Logan, UT) supplemented with 5% fetal bovine serum, 1% a,l-glutamine, 1%OPI, and two mM glutamine in RPMI as previously described7 (link). All cells were incubated in a 5% CO2 atmosphere at 37°C and trypsin-passaged at 80% confluence.
Yavuz B., Zeki J., Taylor J., Harrington K., Coburn J.M., Ikegaki N., Kaplan D.L, & Chiu B. (2019). SILK RESERVOIRS FOR LOCAL DELIVERY OF CISPLATIN FOR NEUROBLASTOMA TREATMENT: IN VITRO AND IN VIVO EVALUATIONS. Journal of pharmaceutical sciences, 108(8), 2748-2755.
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5

Determination of CX-5461 Cytotoxicity in Neuroblastoma Cell Lines

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MYCN-amplified human neuroblastoma cell lines (KELLY, IMR5) and non-MYCN-amplified cell lines (SY5Y, SKNAS) were used in this study. Of note, the non-MYCN-amplified cell lines express high levels of MYC (c-Myc) protein. Both MYCN and MYC proteins belong to the MYC family proteins. KELLY cells were purchased from Sigma-Aldrich (St Louis, MO); IMR5 cells were a gift of Roger H. Kennett (University of Pennsylvania, PA. USA); SY5Y and SKNAS cell lines were obtained from American Type Culture Collection (ATCC, Manassas, VA). All cell lines were maintained in RPMI 1640 (HyClone, Logan, UT) supplemented with 5% fetal bovine serum, 1% OPI, 1% a,l-glutamine, and 250 µL gentamycin. All cells were maintained in a 5% CO2 atmosphere at 37°C and passaged via trypsinization when they reached 80% confluence. In order to determine the half maximal inhibitory concentration (IC50) of CX-5461, all cell lines were exposed to varying doses of CX-5461 for two days. The IC50 was defined as the amount of drug required to kill 50% of the cells.
Taylor J.S., Zeki J., Ornell K., Coburn J., Shimada H., Ikegaki N, & Chiu B. (2019). Down-Regulation of MYCN Protein by CX-5461 Leads to Neuroblastoma Tumor Growth Suppression. Journal of pediatric surgery, 54(6), 1192-1197.
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6

Cell Line Culture and Characterization

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BE(2)C (ATCC #CRL-2268), PA-1 (ATCC #CRL-1572), IMR90 (ATCC #CRL-186), SK-N-AS (ATCC #CRL-2137), SH-SY5Y (ATCC #CRL-2266), 293T(ATCC #11268), SK-N-DZ (ATCC #CRL-2149) and Kelly cells (Sigma #92110411-1VL) were maintained in 1:1 DMEM/F12:MEM media with 10% inactivated fetal calf serum, 1ug/ml penicillin, and 1U/ml streptomycin. All cell lines were purchased for the purposes of this study, are not among commonly misidentified cell lines (per ICLAC), and tested negative for mycoplasma contamination.
Powers J.T., Tsanov K.M., Pearson D.S., Roels F., Spina C.S., Ebright R., Seligson M., de Soysa Y., Cahan P., Theiβen J., Tu H.C., Han A., Kurek K.C., LaPier G.S., Osborne J.K., Ross S.J., Cesana M., Collins J.J., Berthold F, & Daley G.Q. (2016). Multiple mechanisms disrupt the let-7 microRNA family in neuroblastoma. Nature, 535(7611), 246-251.
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7

Culturing Human Neuroblastoma KELLY Cells

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Human neuroblastoma KELLY cells (Sigma-Aldrich, St Louis, MO) were maintained in RPMI 1640 (HyClone, Logan, UT) supplemented with 10% fetal bovine serum, 100 IU/ml penicillin, 100 µg/ml streptomycin, and 2 mM glutamine. All cells were maintained in a 5% CO2 atmosphere at 37°C and trypsin-passaged at 80% confluence.
Coburn J.M., Harris J., Cunningham R., Zeki J., Kaplan D.L, & Chiu B. (2017). Manipulation of variables in local controlled release vincristine treatment in neuroblastoma. Journal of pediatric surgery, 52(12), 2061-2065.
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8

Coimmunoprecipitation of MYCN and EYA1 in KELLY cells

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KELLY cells were purchased from Sigma (St. Louis, MO) and cultured in DMEM:F12, supplemented with 10% FBS. HEK293TN cells were cultured in DMEM with 10% FBS. Transient transfection was performed by using jetPRIME (Polyplus transfection, #114-07) per the manufacturer’s instructions. CoIP analysis was performed essentially as previously described[21 (link)]. Briefly, two days post-transfection, whole cell lysates of HEK293TN cells were prepared and used for V5-IP (Invitrogen, #R960-25). In the case of KELLY cells, nuclear extracts were isolated and subjected to immunoprecipitation with anti-MYCN antibody (Santa Cruz, #sc-53993), followed by immunoblotting with anti-EYA1 antibody (Prosci, #25-067). The following antibodies were used in western blot: α-Flag (Thermo Fisher Scientific, #MA1-91878) and β-actin (Sigma, #A5316).
Hansen J.N., Lotta LT J.r., Eberhardt A., Schor N.F, & Li X. (2016). EYA1 expression and subcellular localization in neuroblastoma and its association with prognostic markers. Journal of cancer research & therapy, 4(2), 11-18.
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9

Culturing Human Neuroblastoma Cell Lines

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Human neuroblastoma cell lines SK-N-AS (American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10 % fetal bovine serum, 100 IU/ml penicillin, 100 μg/ml streptomycin, and 0.1 mM non-essential amino acids. KELLY cells (Sigma-Aldrich, St Louis, MO, USA) were maintained in RPMI 1640 supplemented with 10 %v/v fetal bovine serum, 100 IU/ml penicillin, 100 μg/ml streptomycin, and 2 mM glutamine. All cells were maintained in a humidified 5 % CO2 atmosphere at 37°C and were trypsin-passaged at 80 % confluence.
Seib F.P., Coburn J., Konrad I., Klebanov N., Jones G.T., Blackwood B., Charest A., Kaplan D.L, & Chiu B. (2015). Focal therapy of neuroblastoma using silk films to deliver kinase and chemotherapeutic agents in vivo. Acta biomaterialia, 20, 32-38.
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10

Neuroblastoma Cell Lines: Genetic Diversity

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Four neuroblastoma cell lines with different genetic makeup were used. Human neuroblastoma SK-N-AS cells (American Type Culture Collection (ATCC, Manassas, VA, USA)) were maintained in Dulbecco's modified Eagle's medium (DMEM, HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, 100 μg ml−1 streptomycin, and 0.1 mM non-essential amino acids. Human neuroblastoma IMR-32 cells (ATCC) were maintained in modified Eagle's medium (MEM, HyClone) supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, and 100 μg ml−1 streptomycin. Human neuroblastoma SH-SY5Y cells (ATCC) were maintained in 1 : 1 mixture of MEM and F-12 media supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, and 100 μg ml−1 streptomycin. Human neuroblastoma KELLY cells (Sigma-Aldrich, St Louis, MO, USA) were maintained in RPMI-1640 (HyClone) supplemented with 10% fetal bovine serum, 100 IU ml−1 penicillin, 100 μg ml−1 streptomycin, and 2 mM glutamine. All cells were maintained in a 5% CO2 atmosphere at 37 °C and trypsin-passaged at 80% confluence.
Chiu B., Coburn J., Pilichowska M., Holcroft C., Seib F.P., Charest A, & Kaplan D.L. (2014). Surgery combined with controlled-release doxorubicin silk films as a treatment strategy in an orthotopic neuroblastoma mouse model. British Journal of Cancer, 111(4), 708-715.
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