Alpha TC1 clone 6 (ATCC Cat. No. CRL-2934, RRID:CVCL_B036), referred to as α cells (ATCC, Manassas, VA), were cultured in DMEM 6046 (Sigma-Aldrich) supplemented with 10% (v/v) fetal bovine serum (FBS; Sigma-Aldrich), 15 mM HEPES, 0.1 mM nonessential amino acids (NEAA), 1.5% (wt/vol) sodium bicarbonate, and 2.0% (wt/vol) glucose. INS-1E (RRID:CVCL_0351), referred to as β cells (AddexBio, San Diego, CA), were cultured in RPMI-1640 (Gibco) supplemented with 5% (vol/vol) FBS, 1 mM sodium pyruvate, 10 mM HEPES, and 0.05 mM 2-mercaptoethanol. Human umbilical vein endothelial cells (HUVECs) (C2519A, used at passage 5; Lonza, Walkersville, MD), were cultured in EGM-2 (PromoCell; C-22111). All cell types were negative for mycoplasma contamination (Mycoplasma Detection Kit, #B39035; BioTool) and were cultured under a humidified atmosphere with 5% CO2 at 37°C.
Ins 1e
Manufactured by Addexbio
Sourced in United States
The INS-1E is a cell line commonly used in diabetes research. It is derived from rat insulinoma cells and exhibits characteristics of pancreatic beta cells. The INS-1E line is frequently utilized for in vitro studies related to insulin secretion, beta cell function, and other aspects of glucose metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Dmem 6046, by Merck Group (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Egm 2, by PromoCell (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Interleukin il 1β, by R&D Systems (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Ins 1e, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Harvard Bioscience (1 mentions)
Recombinant sfrp5, by R&D Systems (1 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Chir99021, by Miltenyi Biotec (1 mentions)
Alpha tc1 clone 6, by American Type Culture Collection (1 mentions)
Brin bd11, by Merck Group (1 mentions)
6 protocols using ins 1e
1
Cell Culture Protocols for Islet and Endothelial Cells
2
Culturing Rat Pancreatic Beta Cells
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The rat pancreatic β-cell line INS-1E (AddexBio, San Diego, CA) passage 30–70 was cultured in RPMI-1640 media supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, 10 mM HEPES, 1 mM sodium pyruvate and 50 μM 2-mercaptoethanol. Cells were cultured in a humidified atmosphere at 37 °C with 5% CO2 in air.
3
INS-1E Cell Culture and Treatments
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We seeded 40,000 INS-1E (AddexBio, San Diego, CA, USA) per cm2 (flasks) and cultivated these cells in medium containing RPMI 1640 with glutamine (Life Technologies, Carlsbad, CA, USA), 1 mmol/l sodium pyruvate (Life Technologies), 50 μmol/l β-mercaptoethanol (Life Technologies), 10 mmol/l HEPES (Life Technologies), 10% fetal bovine serum (FBS) (Biochrom, Berlin, Germany), 100 IU/ml penicillin and 100 μg/ml streptomycin (Life Technologies). After 4 days, INS-1E were detached using 0.05% trypsin (Life Technologies), seeded at 100,000 cells per cm2 (24- or 96-well plate) and cultured in the aforementioned medium for another 3 days. Then, cells were fasted for FBS for 4h and treated with 0.1, 1 or 5 μg/ml recombinant Sfrp5 (R&D Systems, Wiesbaden, Germany) for 24h. INS-1E cells were also incubated with 0.2 ng/ml interleukin (IL)-1β (R&D Systems) as positive control for the inhibition of cell viability. Treatment of the cells with 10 μmol/l CHIR99021 (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) served as positive control of an activated canonical Wnt signalling pathway.
4
In Vitro Cell Culture Protocol
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Alpha TC1 clone 6 (Cat# CRL-2934 ATCC, Manassas, United States), referred to as α cells, were cultured in DMEM 6046 (Sigma-Aldrich) supplemented with 10% (vol/vol) FBS (Sigma-Aldrich), 15 mM HEPES, 0.1 mM non-essential amino acids, 1.5% (wt/vol) sodium bicarbonate and 2.0% (wt/vol) glucose. INS-1E (AddexBio, San Diego, United States), referred to as β cells, were cultured in RPMI 1640 (Gibco) supplemented with 1 mM sodium pyruvate, 0.05 mM 2-mercaptoethanol, 10 mM HEPES, and 5% (vol/vol) FBS. Human umbilical vein endothelial cells (HUVECs) (C2519A, Lonza, Maryland, United States; used at passage 5) were cultivated in EGM-2 (PromoCell). All cells were cultured under a humidified atmosphere with 5% CO2 at 37°C and negative for mycoplasma contamination (Mycoplasma Detection Kit, #B39035, BioTool).
5
Mycoplasma-free Cellular Coculture Assay
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INS-1E (AddexBio #C0018009) and C2C12 (ATCC CRL-1772) were commercially available and therefore not authenticated following purchase. The cells were free of mycoplasma as determined by PCR.
6
Cultivation of Clonal β-Cell Lines
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The clonal β-cell line BRIN-BD11 (Sigma-Aldrich, Cat no. 10033003) is a hybrid line formed by electrofusion of a primary culture of rat pancreatic β-cells (New England Deaconess Hospital, NEDH) with RINm5F cells [51 (link),52 (link)]. The cells were cultured in a humidified atmosphere containing 5% CO2 and at 37°C in a complete medium, composed of RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 2 mM l -glutamine, 10 mM HEPES, 100 U ml−1 penicillin and 100 U ml−1 streptomycin.
The rat clonal β-cell line INS-1E [53 (link)] (AddexBio, Cat no. C0018009) derived from parental INS-1 cells [54 (link)] was cultured in a humidified atmosphere containing 5% CO2 and at 37°C in a complete medium, composed of RPMI 1640 medium supplemented with 10% heat-inactivated FBS, 1 mM sodium pyruvate, 50 mM 2-mercaptoethanol, 2 mMl -glutamine, 10 mM HEPES, 100 U ml−1 penicillin and 100 U ml−1 streptomycin.
Mouse insulinoma-derived MIN6 cells were a kind gift from the Miyazaki laboratory, Osaka University, Japan [55 (link)] and were cultivated in a complete medium, composed of DMEM high glucose medium, supplemented with 10% heat-inactivated FBS, 50 mM 2-mercaptoethanol, 2 mMl -glutamine, 100 U ml−1 penicillin and 100 U ml−1 streptomycin.
The rat clonal β-cell line INS-1E [53 (link)] (AddexBio, Cat no. C0018009) derived from parental INS-1 cells [54 (link)] was cultured in a humidified atmosphere containing 5% CO2 and at 37°C in a complete medium, composed of RPMI 1640 medium supplemented with 10% heat-inactivated FBS, 1 mM sodium pyruvate, 50 mM 2-mercaptoethanol, 2 mM
Mouse insulinoma-derived MIN6 cells were a kind gift from the Miyazaki laboratory, Osaka University, Japan [55 (link)] and were cultivated in a complete medium, composed of DMEM high glucose medium, supplemented with 10% heat-inactivated FBS, 50 mM 2-mercaptoethanol, 2 mM
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