Hyclon

All colorectal cancer cell lines (LoVo, SW48, SW480, and SW620) were obtained from the Type Culture Collection cell bank (Chinese Academy of Sciences, Beijing, PR China). Cells were cultured at 37°C, 5% CO₂ in Dulbecco Modified Eagle Medium (DMEM), supplemented with 10% fetal bovine serum (Hyclon; Thermo Scientific, Rockford, IL, USA).

Bone marrow cells (BM cells) were isolated from the femoral bone of healthy Balb/c mice (male, weight 19 ± 1 g). BM cells were suspended in the complete growth medium based on Dulbecco’s modified Eagle’s medium (DMEM) (Sigma-Aldrich, St. Louis, MO, USA), supplemented with 10% fetal bovine serum (FBS; HyClon, Thermo Fisher Scientific, Waltham, Massachusetts, USA), 1% penicillin/streptomycin (PanEco, Moscow, Russia) and 4 mM L-glutamine (PanEco, Moscow, Russia) at 37 °C in atmosphere with 5% CO₂ to a concentration of 500,000 cells/mL. To study cell activity, the suspension of BM cells (150 μL) was placed in E-plates 16 (ACEA Biosciences, San Diego, CA, USA). Solutions of LF, LF-deS, and rG-CSF in isotonic sodium chloride solution (50 μL) were added to the cells to a concentration of 50 µg/mL or 250 µg/mL for polysaccharides and 0.15 nmol/mL or 0.75 nmol/mL for rG-CSF. BM cells supplemented with 50 μL isotonic sodium chloride solution were used as an intact Control. The results were examined daily by assessing the change in the Cell Index in comparison with the Control, which was measured using the Agilent xCELLigence real-time cell analysis multiple plates system (xCELLigence RTCA DP, ACEA Biosciences, San Diego, CA, USA) during incubation for 7 days at 37 °C in atmosphere with 5% CO₂. A fresh portion of the growth medium (50 μL) was added every two days in each well.