Hacat cells

Six components of S. flexibilis including, 11-dehydrosinulariolide (SC-2), sinulariolide (SC-4), dendronpholide F (SC-6), 3,4:8,11-bisepoxy-7-acetoxycembra-15(17)-en-1,12-olide (SC-7), sinularin (SC-9), and dihydrosinularin (SC-10), were isolated and identified by Prof. Jui-Hsin Su (National Museum of Marine Biology and Aquarium, Pintung, Taiwan) [23 (link)]. Dimethyl sulfoxide (DMSO), LPS, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), napthyl-ethylene-dihydrochloride, sulfanilamide, and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA). RAW 264.7 (BCRC 60001) and HaCaT cells were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan). Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), antibiotics, and glutamine were purchased from GIBCO BRL (Grand Island, NY, USA). The bacterial culture equipment, including an anaerobic atmosphere by MGC AnaeroPack-Jar and MGC AnaeroPack-Anaero, were purchased from Mitsubishi Gas Chemical (Tokyo, Japan). Blood agar plates (BAPs), BactoTM tryptic soy agar (TSA), and BactoTM tryptic soy broth (TSB) were purchased from Difco (Detroit, MI, USA).

Ulva lactuca was obtained from Taiwan Fertilizer Co. (Taipei, Taiwan). Chitosan (CS; molecular weight of 192 kDa, degree of deacetylation of ca. 85%) was supplied by Sigma Chemical Co. (St. Louis, MO, USA). Murine RAW 264.7 (murine macrophage), NIH 3T3 (mouse fibroblast), and HaCaT cells (human keratinocyte cells) were provided by the Bioresource Collection and Research Center (BCRC), Hsinchu, Taiwan. The culture medium for NIH 3T3 and HaCaT cells was DMEM/High glucose medium (Gibco^®, Thermo Fisher Scientific Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS), 3.7 g of sodium bicarbonate and antibiotics (100 U/mL Penicillin and 100 μg/mL Streptomycin), whereas the medium for culturing RAW 264.7 was also DMEM/High glucose medium yet supplemented with 10% FBS, 2.5 g of sodium bicarbonate, 3.7 g of HEPES and antibiotics (100 U/mL of Penicillin and 100 μg/mL of Streptomycin). All other chemicals were of reagent grade and purchased from Sigma Chemical Co., unless stated otherwise.

Hs68, HaCaT cells, and B16F0 cells were purchased from the Bioresource Collection and Research Center in Hsinchu, Taiwan. These cells were maintained in DMEM containing 10% FBS, 100 U/mL penicillin, and 100 U/mL streptomycin in an incubator set at 37 °C.