Authenticated human NB cell lines were obtained from certified suppliers between 2014 and 2016: SK-N-FI from the Childhood Cancer Repository/Children’s Oncology Group Resource Laboratory, SK-N-AS and SK-N-BE from the ATCC, and Kelly from Sigma-Aldrich. The SK-N-BE clone used was SK-N-BE(2)c (ATCC: CRL-2268 ). All cell lines were maintained in RPMI medium (Corning) supplemented with 10 % heat-inactivated fetal bovine serum (Hyclone), penicillin (100 IU/ml), and streptomycin (100 μg/ml). Cell lines were routinely monitored for mycoplasma contamination every six months using the MycoAlert™ PLUS mycoplasma detection kit (Lonza).
Sk n be
Manufactured by American Type Culture Collection
Sourced in United States, Italy
SK-N-BE is a human cell line derived from a neuroblastoma. It is commonly used in research and laboratory settings for various applications.
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Lab products found in correlation
Sk n sh, by American Type Culture Collection (16 mentions)
Imr 32, by American Type Culture Collection (14 mentions)
Sh sy5y, by American Type Culture Collection (13 mentions)
Sk n as, by American Type Culture Collection (12 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Streptomycin, by Thermo Fisher Scientific (7 mentions)
Penicillin, by Thermo Fisher Scientific (7 mentions)
Sk n dz, by American Type Culture Collection (6 mentions)
Sh sy5y, by Thermo Fisher Scientific (5 mentions)
Sk n be, by Thermo Fisher Scientific (5 mentions)
Sk n sh, by Thermo Fisher Scientific (4 mentions)
Chp134, by American Type Culture Collection (4 mentions)
Sms san, by American Type Culture Collection (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (3 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
Imr 32, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Nbl s, by American Type Culture Collection (3 mentions)
37 protocols using sk n be
1
Neuroblastoma Cell Line Authentication and Maintenance
2
Evaluating SPIO NPs and ASOs in Neuroblastoma
3
Detailed Cell Line Acquisition and Culture
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The human MM cell line MM.1S was provided by Dr ST Rosen (Northwestern University, Chicago, IL, USA), whereas the MM.1S-luc cell line was donated by Dr CS Mitsiades (Dana-Farber Cancer Institute, Boston, MA, USA). RPMI8226 cells purchased from the American Type Culture Collection (ATCC) were lentivirally transduced to stably express firefly luciferase as in Groen et al. [80 (link)]. Other established MM cell lines (OPM-2, JJN3), together with the erythroleukemia (HEL) and chronic B lymphocytic leukemia (MEC-1) cell lines were purchased from the Leibniz Institute DSMZ biosource center. All hematological tumor cell lines were periodically authentified by STR DNA profiling, and grown in RPMI 1640 medium or IMDM (MEC-1) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine.
The BM endothelial cell line BMEC-1 was provided by Dr FJ Candal (Centers for Disease Control and Prevention, Atlanta, Georgia, USA). This cell line was grown in MCDB 131 medium containing 15% FBS, 2.5 μg/ml amphotericin B, 1 μg/ml hydrocortisone, 10 ng/ml EGF and antibiotics. The human neuroblastoma cell line SK-N-BE was purchased from ATCC and grown in DMEM medium (4.5 g/l glucose) with 10% FBS and antibiotics.
The BM endothelial cell line BMEC-1 was provided by Dr FJ Candal (Centers for Disease Control and Prevention, Atlanta, Georgia, USA). This cell line was grown in MCDB 131 medium containing 15% FBS, 2.5 μg/ml amphotericin B, 1 μg/ml hydrocortisone, 10 ng/ml EGF and antibiotics. The human neuroblastoma cell line SK-N-BE was purchased from ATCC and grown in DMEM medium (4.5 g/l glucose) with 10% FBS and antibiotics.
4
Culturing Panc-1 and SK-N-BE Cell Lines
5
Culturing Human Neuroblastoma Cell Lines
6
Neuroblastoma Cell Line Response to Oxidative Stress
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Human neuroblastoma cell line, SK-N-BE(2) (CRL-2271, ATCC®, LGC Standards S.r.l., Milan Italy) were cultured at 37 °C in a 5% CO2 humidified incubator in either RPMI-1640 medium (Euroclone spa, 20016 Pero, MI) supplemented with 10% fetal bovine serum (FBS), glutamine (2 mM), sodium pyruvate and antibiotics (0.02 mg/mL streptomycin and 0.02 IU/mL penicillin).
Ginkgo biloba L. extract EGb 761 (EGb) was a gift from Schwabe (Schwabe Pharma Italia Srl, Egna, Italy). EGb stock solution contained 250 mg/mL of extract was dissolved in dimethyl sulfoxide (DMSO). Hydrogen peroxide (H2O2,) (Sigma-Aldrich, St. Louis, MI, USA) was used as oxygen stress inducer.
Ginkgo biloba L. extract EGb 761 (EGb) was a gift from Schwabe (Schwabe Pharma Italia Srl, Egna, Italy). EGb stock solution contained 250 mg/mL of extract was dissolved in dimethyl sulfoxide (DMSO). Hydrogen peroxide (H2O2,) (Sigma-Aldrich, St. Louis, MI, USA) was used as oxygen stress inducer.
7
Cell Line Cultivation and Maintenance
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The human NB cell line IMR32 (ATCC: CCL-127), the NB cell line SKN-BE (ATCC: CRL-2271), and the human glioma cell lines U87MG (ATCC: HTB-14), T98G (ATCC: CRL-1690), and A172 (ATCC: CRL-1620) were all purchased from ATCC (Manassas, VA, USA). The human acute myelocytic leukemia cell line Molm-13, human epidermal cancer cell line A431, and human breast cancer cell line MDA-MB-231 were all obtained from Cell Resource Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/Peking Union Medical College (IBMS, CAMS/PUMC). Adherent cells were cultured in DMEM, and cells were cultured in suspension with RPMI 1640 medium with 10% FBS and a mixture of penicillin/streptomycin. Cells were cultured at 37°C in a humidified atmosphere with 5% CO2. All experiments were performed on cells in the exponential growth phase.
8
Cell Culture Conditions for Receptor Studies
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SK-N-BE and MCF-7 cell lines (ATCC, LGC Standards S.r.l., Milan, Italy) were used at passage 4–8 and were grown in air containing 5% CO2 in phenol red free, RPMI-1640 or DMEM medium, respectively, containing 10% (v/v) charcoal-stripped fetal bovine serum, L-glutamine (2.0 mM), Pen-Strep solution (penicillin 100 U/ml and streptomycin (100 mg/ml) as previously described [8 ,24 (link)] (Control Medium). Nutrient deprivation condition was obtained by culturing cells in amino acid and serum free, Earle’s Balanced Salt Solution (EBSS, Sigma Aldrich) containing 1 g/L of D-glucose for the indicated times. Stable ERα-transfected HEK-293 (ERα-HEK-293) cell lines were routinely grown in media containing G418 50 mM [25 (link)]. Cell line authentication were periodically performed by amplification of multiple STR loci by BMR Genomics srl (Padua, Italy). Cells were simultaneously treated with the vehicle used to dissolve all drugs (ethanol/PBS 1:10, v/v), and/or E2 (1 or 10 nM), and/or Bafalomycin-A1 (Baf-A1, 100 nM). When indicated, E2 or Baf-A1 pretreatment were performed adding the compounds 1 h before. For nutrient deprivation, MCF-7, SK-N-BE or ERα-HEK-293, were cultured as above reported, washed 3 times with PBS then cultured in EBSS for the indicated time points.
9
Culturing Human Neuroblastoma Cell Lines
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The human NB cell lines KELLY (also known as N206; #92110411 from Sigma), SK-N-AS (called SKNAS in this study; CRL-213 from ATCC), SK-N-BE (clone SKNBE(2)C, called SKNBE in this study; CRL-2268 from ATCC), and SK-N-FI (called SKNFI in this study; from the Children’s Oncology Group) were cultured in RPMI 1640 (VWR), supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen), Penicillin (100 IU/mL) and Streptomycin (100 μg/mL) (30-002-CI, Corning). All cells were purchased from their respective suppliers within the last 2 years. The cells were maintained at 37 °C in a humidified atmosphere containing 5% CO2.
10
Establishing SIRT4 Overexpression Cell Lines
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Human NB cell lines SH-SY5Y (CRL-2266), SK-N-BE (CRL-2271), IMR-32 (CCL-127), and 293T (CRL-3216) were purchased from ATCC (Manassas, VA, USA). Cells were cultured in DMEM (D6046; Sigma-Aldrich Co., St Louis, MO, USA) containing 10% FBS (10099–141; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) at 37°C and 5% CO2. No antibiotics were added to the growth medium. Cell lines overexpressing SIRT4 were established by transfection with pLenti-C-Myc-DDK-IRES-Puro Tagged Cloning Vector (PS100069; ORIGEN, Annapolis, MD, USA) and Lentiviral Packaging Kits (TR30037; ORIGEN) according to the manufacturer’s protocol. Cells transfected with empty plasmids were used as control. Puromycin at the concentration of 2 mg/mL for SH-SY5Y, 2 mg/mL for SK-N-BE and 3 mg/mL for IMR-31 to select the stable cell expression, and cell lines with SIRT4 overexpression or knockout were established.
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