The B16-F10 mouse melanocyte cell line was kindly provided by Dr. Benilde Jiménez Cuenca, Instituto de Investigaciones Biomédicas “Alberto Sols” UAM-CSIC. Human dermal fibroblasts (HDF) were obtained from a skin biopsy by trypsinization with 0.25% Trypsin-Ethylenediaminetetraacetic acid (EDTA). Cells were cultured in Dulbecco’s modified eagle medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS), 1% (v/v) penicillin G (100 U/mL), and streptomycin (100 µg/mL) (HyClone Laboratories, South Logan, UT, USA). Cells were maintained under standard conditions at 37 °C, 5% humidity, and 5% CO2 in an incubator (Heraeus HERAcell, Thermo Scientific, Waltham, MA, USA).
Heraeus heracell
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Heraeus HERAcell is a reliable and efficient incubator designed for cell culture applications. It offers precise temperature and CO2 control to maintain optimal growth conditions for cell lines. The incubator features a durable stainless-steel interior and advanced technology to ensure consistent environmental parameters.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Cytiva (5 mentions)
Penicillin g, by Cytiva (4 mentions)
Fetal bovine serum (fbs), by Cytiva (2 mentions)
Penicillin, by Cytiva (1 mentions)
Low glucose dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Vero cell line atcc ccl81, by American Type Culture Collection (1 mentions)
Cercopithecus aethiops, by American Type Culture Collection (1 mentions)
6 protocols using heraeus heracell
1
Culturing B16-F10 Melanocytes and Human Dermal Fibroblasts
2
Culturing Murine Melanoma Cells for Research
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The B16-F10 mouse melanocyte cell line was kindly provided by Dr. Benilde Jiménez Cuenca, Instituto de Investigaciones Biomédicas «Alberto Sols» UAM-CSIC (Madrid, Spain). Cells were cultured in Dulbecco’s modified eagle medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS), 1% (v/v) penicillin G (100 U/mL) and streptomycin (100 µg/mL) (HyClone Laboratories, South Logan, UT, USA). Cells were maintained under standard conditions at 37 °C, 5% humidity and 5% CO2 in an incubator (Heraeus HERAcell, Thermo Scientific, Waltham, MA, USA). B16-F10 cell line is a melanoma cell line and not normal melanocytes, but we consider that this cell line is appropriate for the present study. We consider that the use of a malignant cell line does not interfere with the parameters that have been evaluated in the study.
3
Human Dermal Fibroblast and Keratinocyte Culture
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Human dermal fibroblasts (HDF) were obtained from a skin biopsy after two rounds of trypsinization with 0.25% Trypsin-EDTA by shaking at 37° C. We also used the keratinocyte cell line HaCaT. Cells were cultured in Dulbecco’s modified eagle medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) and penicillin (50 μg/mL) and streptomycin (50 μg/mL) (HyClone Laboratories, South Logan, UT, USA). Cells were maintained at 37 °C, 95% humidity and 5% CO2 in an incubator (Heraeus HERAcell, Thermo Scientific, Waltham, MA, USA).
4
Isolation and Culture of Human Dermal Fibroblasts
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Human dermal fibroblasts (HDF) were obtained from a skin biopsy after two rounds of trypsinization with 0.25% Trypsin-EDTA by shaking and at 37 °C. HDF were cultured in Dulbecco’s modified eagle medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% (v/v) penicillin G (100 U/mL) and streptomycin (100 µg/mL) (HyClone Laboratories, South Logan, UT, USA). Cells were maintained at 37 °C, 5% humidity and 5% CO2 in an incubator (Heraeus HERAcell, Thermo Scientific, Waltham, MA, USA).
5
Cell Culture of Huh-7 and Vero Cells
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The Huh-7 hepatocarcinoma cell line was generously provided by Dr. Sonia Zúñiga (CNB-Spanish National Centre for Biotechnology, Madrid, Spain). This cell line was obtained from a liver tumor in a 57-year-old Japanese male and was used to test HCoV-229E infection [33 (link)]. The Vero cell line (ATCC-CCL81), which was derived from the kidney of an adult African green monkey (Cercopithecus aethiops) was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). All cell lines were cultured in growth medium (GM) containing low-glucose Dulbecco’s modified Eagle medium (Life Technologies, Paisley, UK) supplemented with 5% fetal bovine serum (FBS), 1% (v/v) penicillin G (50 U/mL) and streptomycin (50 µg/mL) (HyClone Laboratories, Logan, UT, USA). Cells were maintained at 37 °C, in a humidified atmosphere of 5% CO2 (Heraeus HERAcell, Thermofisher. Waltham, MA, USA).
6
Isolation and Culture of Human Dermal Fibroblasts and Mouse Melanocytes
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HDF were obtained from a skin biopsy after trypsinization with 0.25% Trypsin-EDTA. B16-F10 mouse melanocyte cell line was kindly provided by Dr. Benilde Jiménez Cuenca, Instituto de Investigaciones Biomédicas «Alberto Sols» UAM-CSIC. Both HDF and melanocytes were cultured in Dulbecco’s modified eagle medium (DMEM) supplemented with 1% (v/v) penicillin G (100 U/mL) and streptomycin (100 µg/mL) and 10% (v/v) fetal bovine serum (FBS) (HyClone Laboratories, South Logan, UT, USA). Cells were maintained at 37 °C, 5% humidity and 5% CO2 in an incubator (Heraeus HERAcell, Thermo Scientific, Waltham, MA, USA).
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