, Ltd.), Mitomycin (MMC), Adriamycin (ADR) (MMC and ADR RG7420 cost obtained from Zhejiang Hisun Pharmaceutical Co., Ltd.), Vincristine (VCR), Paclitaxel (PTX) (VCR and PTX obtained from Shanghai Hualian Pharmaceutical Factory) and 5-flurouracil (5-FU) (Shanghai Xudong Pharmaceutical Selleck A-1210477 Co., Ltd.). Effector cells Preparation and in vitro amplification of CIK cells: The periphery heparin from healthy adults was obtained for anticoagulation, and prepared according to a previous report by Schmidt-Wolf
IG et al. [17], cells were harvested in the 14th day, and the ratio of potency and target was adjusted to 40:1, 20:1 or 10:1 before use. Construction and grouping of the human gastric cancer OCUM-2MD3/L-OHP cell peritoneal transplantation model Preliminary experiments using our assay confirmed that the incidence of peritoneal tumors was 100% when each Balb/c nude mouse (female, 4~6 week, 15~18 g, animal licenses lot: SCXK 11-00-0005) was inoculated intraperitoneally with 5 × 106 drug-resistant cells. In our experiment, 35 nude mice were selected and inoculated intraperitoneally with drug-resistant cells at a dose of 5 × 106 cells per 0.2 ml each, and the human XAV-939 solubility dmso gastric cancer drug resistant cell peritoneal transplantation model was established. All mice were randomly divided
into seven groups, including the normal control, NS control, L-OHP (1.125 mg/kg, 2.25 mg/kg), CIK (2 × 107/0.2 mL, 4 × 107/0.2 mL) and L-OHP+CIK groups. Intraperitoneal injection of drug-resistant cells was performed in the first six groups after 15 days of inoculation, once every other day for a total of three injection days. L-OHP (1.125 mg/kg) was administered to the L-OHP+CIK group after inoculation Thalidomide for 15 days, then CIK cells (2 × 107/0.2 mL/number) were injected intraperitoneally twice every other day for a total of three injection days. Methods Observation of cell biological characteristics of OCUM-2MD3/L-OHP (Parental cells were used as control)
Cell morphology observation of drug-resistant cells Both cell types were cultured on culture plates and observed under an inverted phase contrast microscope until the cells covered 80% of the bottom wall. Cells were collected (1 × 107 ), fixed with 2.5% glutaraldehyde followed by 2% osmium tetroxide, dehydrated, embedded, sectioned, stained and observed and photographed with a transmission electron microscope. Growth curve of OCUM-2MD3/L-OHP cells by cell count method The two cell types were inoculated into 24-well plates at a density of 1.5 × 104 cells/well and cultured at 37°C in a humidified incubator containing 5% CO2. Three wells were used for live-cell counts each day, and a cell-growth curve was plotted after counting cells continuously for six days.