The human keratinocyte (HaCaT) cell line was purchased from Procell Life Science & Technology (CL-0090) and cultured with Minimum Essential Medium (MEM; Procell Life Science & Technology, PM150410) containing 10% fetal bovine serum (Gibco, A3160802). Normal human epithelial keratinocytes (NHEK) were purchased from Beijing Beina Chuanglian Biotechnology Institute (BNCC340593) and cultured with Minimum Essential Medium with Earle’s Balanced Salts (MEM/EBBS; Hyclone, SH30024.01) containing 10% fetal bovine serum (Gibco, A3160802). Cells were incubated in humidified condition of 37°C with 95% air and 5% CO2. For experiments, HaCaT cells or NHEK were treated with different concentrations of glucose (Sigma-Aldrich, G6152; normal glucose: 5.6 mM of glucose; mid-high glucose: 15 mM of glucose; high glucose: 30 mM glucose). mannitol (Sigma-Aldrich, M4125; 15 mM: 5.6 mM of glucose + 9.4 mM of mannitol; 30 mM: 5.6 mM of glucose + 24.4 mM of mannitol) was used as the osmotic control for mid-high glucose and high glucose.
Cl 0090
Manufactured by Procell
Sourced in China
CL-0090 is a laboratory centrifuge designed for general-purpose applications. It is capable of separating components of a liquid mixture based on their differences in density and size. The centrifuge provides reliable and consistent performance for a variety of sample types and volumes.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Mannitol, by Merck Group (1 mentions)
M4125, by Merck Group (1 mentions)
Sh30024, by Thermo Fisher Scientific (1 mentions)
G6152, by Merck Group (1 mentions)
Pm150410, by Thermo Fisher Scientific (1 mentions)
A3160802, by Thermo Fisher Scientific (1 mentions)
Sp600125, by Selleck Chemicals (1 mentions)
Pm150410, by Procell (1 mentions)
Lipopolysaccharides (lps), by Santa Cruz Biotechnology (1 mentions)
Lipopolysaccharides (lps), by Solarbio (1 mentions)
Minimum essential medium (mem), by Procell (1 mentions)
3 protocols using cl 0090
1
Glucose and Osmotic Stress on Keratinocytes
2
HaCaT Cell Culture and Stimulation
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The HaCaT cells (Procell, CL-0090, Wuhan, China) were obtained from Procell in Wuhan, China, and cultured in MEM medium (Procell, PM150410, Wuhan, China) supplemented with 10% FBS and 1% penicillin/streptomycin at 37 °C in a humidified atmosphere of 5% CO2. Before reagent treatment, the cells were cultured in MEM medium for starvation overnight. The cells grew to 80% confluence before they were stimulated with a specified dose of heat-inactivated C. acnes (1.0 × 107 CFU/mL), FGF21 (20 ng/mL, dissolved in PBS and produced by Wenzhou Medical University Gene Engineering Laboratory, Wenzhou, China), or different inhibitors (10 mM Bay11082, MCE; 20 mM SP600125, Selleck; 10 mM SB203580, MCE, NJ, USA). Then the cells were collected at specified times for RNA isolation, Western blotting, or immunofluorescence and the supernatants were harvested for an enzyme-linked immunosorbent assay (ELISA, Beyotime Biotechnology, PF313).
3
Resveratrol Mitigates LPS-Induced Keratinocyte Injury
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The human immortalized keratinocyte HaCaT cell line (CL-0090; Procell, Wuhan, China) was grown in minimum Eagle's medium (MEM; Procell) with 15% fetal bovine serum (Thermo Fisher, Waltham, MA, USA) and 1% penicillin/streptomycin (Thermo Fisher) in 5% CO 2 at 37 °C.
To mimic skin wounds in vitro, HaCaT cells were stimulated with various doses (0, 2, 4, 6, 8 and 10 μg/mL) of LPS (Solarbio, Beijing, China) for 6 h, as previously reported [24] . To analyze the function of RES in LPS-induced HaCaT cell injury, the cells were exposed to various doses (0, 2, 5 and 10 μM) of RES (C 14 H 12 O 3 ; Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 24 h and then treated with 6 μg/mL LPS for 6 h.
To mimic skin wounds in vitro, HaCaT cells were stimulated with various doses (0, 2, 4, 6, 8 and 10 μg/mL) of LPS (Solarbio, Beijing, China) for 6 h, as previously reported [24] . To analyze the function of RES in LPS-induced HaCaT cell injury, the cells were exposed to various doses (0, 2, 5 and 10 μM) of RES (C 14 H 12 O 3 ; Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 24 h and then treated with 6 μg/mL LPS for 6 h.
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