Ccd 966sk

Elastin Collagen Peptide Complex Juice Drink (YAMII, Zhejiang Kazman Biotechnology Co., China; ingredients: water, fish collagen peptide powder (12%), banana powder, apple essence, cherry essence, γ-aminobutyric acid, grape powder, alma powder, acai powder, perilla seed powder, elderberry powder, brown rice powder, honey, vitamin C, erythritol, citric acid, fructose syrup, sucrose, flavor), human skin fibroblast (CCD-966Sk; ATCC®, CRL-1881), minimum essential medium with 10% FBS, 1% penicillin/streptomycin, and 1 mM sodium pyruvate (Gibco), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Amresco), phosphate-buffered saline (PBS, Gibco), DMSO (ECHO), collagen assay kit (Sircol), mitochondrial membrane potential detection kit (BD), 2′,7′-dichlorofluorescin diacetate (DCFH-DA, Sigma-Aldrich), elastin assay kit (Biocolor), RNA extraction kit (Genaid Biotech), nCounter® platform (NanoString Technologies), flow cytometry (BD) ELISA reader (BioTek).

For testing the proliferation effect of the S. mukorossi seed oil on skin cells, a cell viability assay was performed according to a previous study [36 (link)]. The human skin fibroblast cell line CCD-966SK (ATCC CRL-1881) was used for this in vitro cell analysis. The cells were seeded in 24-well plates at a concentration of 2 × 10⁴ cells/mL and were maintained in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. The cells were then incubated in an environment of 5% CO₂ at 37 °C and 100% humidity. The viability of the CCD-966SK cells exposed to S. mukorossi seed oil at a concentration of 200 μg/mL was evaluated after three days of incubation. The cell viability was assessed using the tetrazolium salt (MTT) method. After the cells were incubated with the tetrazolium salt for 4 h, 500 μL of DMSO were added to solubilize the formazan dye overnight. Since no toxic effects were observed on the cells, the emulsifier polyoxyethylene sorbitan mono-oleate (Tween 80) was employed as the delivery vehicle [38 (link)]. The optical density was determined using a microplate reader (EZ Read 400, Biochrom, Holliston, MA, USA) at 570 and 690 nm.