Reporter cells (293A-TEAD-LUC (HEK-293A-8xGTII-Luc)) were a gift from the laboratory of Xu Wu (Mass General). HEK293A cells were from ThermoFisher. MDCK and HEK293T cells were obtained from the American Type Culture Collection (ATCC). HaCaT immortalized human keratinocytes were from AddexBio. All cell lines were cultured in DMEM (Corning) supplemented with 10% FBS (Gibco) and 1% penicillin/streptomycin (Gibco) and maintained at 37°C with 5% CO2.
Immortalized human keratinocytes hacat
Manufactured by Addexbio
Sourced in United States
Immortalized Human Keratinocytes (HaCaT) are an established cell line derived from normal human skin keratinocytes. The cells have been immortalized through a spontaneous process, retaining their ability to proliferate indefinitely while maintaining their keratinocyte phenotype. HaCaT cells are a widely used model system for the study of various aspects of keratinocyte biology and skin physiology.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Hek293a cells, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle medium dmem, by Merck Group (2 mentions)
Milli q water purification system, by Merck Group (1 mentions)
Adenosine, by Merck Group (1 mentions)
Cordycepin, by Merck Group (1 mentions)
Dulbecco s modified eagle medium, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
5 protocols using immortalized human keratinocytes hacat
1
Cell Line Cultivation Protocols
2
Cell Culture Conditions Specification
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Reporter cells (293A-TEAD-LUC (HEK-293A-8xGTII-Luc)) were a gift from the laboratory of Xu Wu (Mass General). HEK293A cells were from Thermo Fisher, Waltham, MA. MDCK and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA). HaCaT immortalized human keratinocytes were from AddexBio. All cell lines were cultured in DMEM (Corning, Corning, NY) supplemented with 10% FBS (Gibco, Grand Island, NY) and 1% penicillin/streptomycin (Gibco) and maintained at 37 °C with 5% CO2.
3
Formulation and Evaluation of Topical Cordycepin-Based Skincare
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All chemicals, reagents, and standards used were analytical or LC-MS-grade reagents. The water was treated in a Milli-Q water purification system (Millipore, Bedford, Burlington, MA, USA) before use. Sunflower oil was purchased in a local market. Cordycepin (purity ≥ 95.0% HPLC) and adenosine (purity ≥ 99% HPLC) were purchased from Sigma-Aldrich (Milan, Italy). For the o/w topical formulation, cosmetic-grade ingredients were used, such as Carbomer, Sodium Gluconate, Glyceryl Stearate, PEG-100 Stearate, Caprylic/Capric Triglycerides, Cetearyl Alcohol, Cetyl Ricinoleate, Phenoxyethanol, and Ethylhexylglycerin. All listed excipients were purchased from ACEF Spa (Fiorenzuola D’arda, Italy).
Immortalized human keratinocytes (HaCaT) bought from Addexbio Technologies (San Diego, CA, USA) were preserved in Dulbecco’s Modified Eagle Medium (DMEM; Sigma Aldrich, St. Louis, MO, USA) that was supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) in 95% air, 5% CO2, and a humidified atmosphere at 37 °C. Human dermal fibroblasts (HDFs) were preserved in Dulbecco’s Modified Eagle Medium (DMEM; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% of fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) in 95% air, 5% CO2, and a humidified atmosphere at 37 °C.
Immortalized human keratinocytes (HaCaT) bought from Addexbio Technologies (San Diego, CA, USA) were preserved in Dulbecco’s Modified Eagle Medium (DMEM; Sigma Aldrich, St. Louis, MO, USA) that was supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) in 95% air, 5% CO2, and a humidified atmosphere at 37 °C. Human dermal fibroblasts (HDFs) were preserved in Dulbecco’s Modified Eagle Medium (DMEM; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% of fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) in 95% air, 5% CO2, and a humidified atmosphere at 37 °C.
4
Culturing Immortalized Human Keratinocytes and Fibroblasts
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Immortalized Human Keratinocytes (HaCaT), bought from Addexbio Technologies (San Diego, CA, USA), were preserved in Dulbecco’s Modified Eagle Medium (DMEM; Sigma Aldrich, St. Louis, MO, USA) that was supplemented with 10% fetal bovine serum (FBS; Sigma Aldrich, St. Louis, MO, USA) in 95% air, 5% CO2, and humidified atmosphere at 37 °C. Human dermal fibroblasts (HDF) were preserved in Dulbecco’s Modified Eagle Medium (DMEM; Sigma–Aldrich, St. Louis, MO, USA) supplemented with 10% of fetal bovine serum (FBS; Sigma–Aldrich, St. Louis, MO, USA) in 95% air, 5% CO2, and humidified atmosphere at 37 °C. Skin explants, obtained from the skin of healthy female donors (aged 31 and 40) at the surgery center Villa Cinzia (Naples, Italy), were cultured in 24-transwell plates in DMEM/FBS plus antibiotics in air–liquid conditions at 37 °C in 5% CO2 humidified air. All donors had given their written informed consent for the use of the skin tissues, according to the Declaration of Helsinki.
5
Immortalized Keratinocyte and Skin Explant Cultures
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Immortalized Human Keratinocytes (HaCaT), purchased from Addexbio Technologies (San Diego, CA, USA), were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Sigma Aldrich, St. Louis, MO, USA) that was supplemented with 10% fetal bovine serum (FBS; Sigma Aldrich, St. Louis, MO, USA) in 95% air, 5% CO2, and humidified atmosphere at 37 °C.
Skin explants were obtained from the skin of healthy female donors (aged 35–49) following mastectomy or breast reduction procedures at the Villa Cinzia surgery center (Naples, IT). The use of skin tissue was done according to the Declaration of Helsinki and all patients had given their written informed consent. Skin punch biopsies (8 mm) were obtained from skin explants and cultured in 24-transwell plates in DMEM/FBS plus antibiotics in air-liquid conditions at 37 °C in 5% CO2 humidified air.
Skin explants were obtained from the skin of healthy female donors (aged 35–49) following mastectomy or breast reduction procedures at the Villa Cinzia surgery center (Naples, IT). The use of skin tissue was done according to the Declaration of Helsinki and all patients had given their written informed consent. Skin punch biopsies (8 mm) were obtained from skin explants and cultured in 24-transwell plates in DMEM/FBS plus antibiotics in air-liquid conditions at 37 °C in 5% CO2 humidified air.
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