Caco2 cells atcc htb 37

Sea grape algae were cultivated under controlled conditions at Family Farm, Ban Laem District, Phetchaburi, Thailand (N 13° 01′ 37.2”; E 100° 04′ 42.8″), and harvested in August 2019. After collection, the algae underwent rinsing in seawater and brushing to eliminate macroscopic epiphytes and sand particles. Further cleansing with tap water removed residual salt, followed by air drying for 4 days at 25 °C and thermostat drying for 12 h at 60 °C. The dried algae were manually crushed, powdered using a grinder, and the powder's moisture content determined to be 12–15 % by dry weight. The powder was stored in dark bags to avoid light and moisture. The human intestinal epithelial cell line, Caco-2 cells (ATCC® HTB-37™), provided by the American Type Culture Collection (ATCC, Manassas, VA, USA), was cultured in Eagle's Minimum Essential Medium (EMEM) supplemented with l-glutamine and 10 % FBS (fetal bovine serum) from Gibco (Rockville, MD, USA). These cells were incubated under 5 % carbon dioxide at 37 °C in a humidified chamber.

The lingzhi mushrooms (G. lucidum) were supplied by AU Farm, Khlong Toei District, Bangkok, Thailand. The powder from the lingzhi mushrooms was produced using a slightly modified version of the technique proposed by Ketprayoon et al. [25 (link)] Initially, the mushrooms were dried at 60 °C in a hot air oven. A grinder was then used to make a fine powder which was then sieved using a 90 μm mesh. The surface of the mushroom powder then acts to support the hydrolyzing protein. This powder was then stored in a vacuum-sealed polypropylene bag in a desiccator at room temperature until required for further use.
American Type Culture Collection (ATCC, Manassas, VA, USA) provided the human intestinal epithelial cell line, Caco-2 cells (ATCC® HTB-37™) which then underwent culturing in Eagle's Minimum Essential Medium (EMEM) which was supplemented with L-glutamine and 10% fetal bovine serum supplied by Gibco (Rockville, MD, USA). American Type Culture Collection (ATCC, Manassas, VA, USA) also supplied the RAW 264.7 cell lines (ATCC® TIB-71™) which were then grown in Dulbecco's Modified Eagle’s Medium (DMEM) and supplemented using high glucose 4.5 g/L with sodium pyruvate, L-glutamine, and 3.7 g/L NaHCO3 (PAN Biotech, Aidenbach, Bavaria, Germany). The cells were stored until use in a humidified chamber under 5% carbon dioxide at a temperature of 37 °C.