Antibiotic antimycotic solution a002

The mouse macrophage RAW 264.7 cell line was obtained from the Taiwan Bioresource Collection and Research Center (BCRC Number: 60001). Cells were maintained in 90% Dulbecco’s modified Eagle’s medium containing 4 mM L-glutamine, adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose + 10% heat-inactivated fetal bovine serum with 1% Antibiotic Antimycotic Solution (A002, HiMedia Laboratories, India), in a humid field atmosphere of 5% CO₂ at 37 °C.
To examine the toxicity of Sarcodia suieae acetyl-xylogalactan on the RAW 264.7 macrophages, 1 × 10⁶ RAW 264.7 macrophages were treated without (control) and with Sarcodia suieae acetyl-xylogalactan at concentrations of 10 µg/mL, 20 µg/mL and 30 µg/mL for 12 and 24 h at 41 °C under 5% CO₂. At the end of this treatment, a Cell Counting Kit-8 (CCk-8) (B34302, bimake) was used to examine cell survival with a microplate reader at OD 450 nm.

A previous study showed that J774A.1 macrophages are a good model to investigate macrophage polarization (20 (link)). Hence, we selected the J774A.1 cell line to use in this study. The mouse macrophage J774A.1 cell line was obtained from the Taiwan Bioresource Collection and Research Center (BCRC number 60140). Cells were maintained in 90% Dulbecco’s modified Eagle’s medium containing 4 mM L-glutamine, adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose + 10% (v/v) heat-inactivated fetal bovine serum with 1% (v/v) Antibiotic Antimycotic Solution (A002, HiMedia), in a humid field atmosphere of 5% CO₂, at 37°C.