IML的细胞毒性实验分析本研究应用的肺癌细胞在DMEM完全培养液中常规

IML cytotoxicity experiment analysis <br>The lung cancer cells used in this study were routinely cultured in DMEM complete culture medium, which contained 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. The culture conditions were 37°C and 5% CO2 in a humid state. The amount of culture fluid: 35mm petri dish is 2ml culture fluid, 60mm petri dish is 3ml culture fluid, and 10 cm petri dish is 8ml culture fluid. <br>Cryopreservation and recovery of cells: Digest the cells with an appropriate amount of 1.25% trypsin. When the cells are rounded and not yet floated, add an appropriate amount of complete culture medium and repeatedly pipette to form a cell suspension. Mix according to the ratio of cell suspension: glycerol=900ul:100ul, store at -70°C overnight, and store in liquid nitrogen. When resuscitating the cells, quickly thaw the cells in a 37°C water bath and add an appropriate amount of complete culture medium. After 5-8 hours, replace the complete medium and routinely subculture.

Experimental analysis of cytotoxicity in IML<br>Lung cancer cells used in this study were routinely cultured in DMEM full culture fluid, which contained 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. The culture conditions are 37 degrees C and 5% CO2 in a humid state. Amount of culture fluid: 35mm Petri dish for 2 ml of culture liquid, 60 mm Petri dish for 3 ml of culture liquid, 10 cm Petri dish for 8 ml of culture solution.<br>Cell freeze and resuscitation: the cells are digested with the appropriate amount of 1.25% pancrease, when the cells become round, not yet floating when added a suitable amount of complete culture fluid and repeated blowing to form cell suspension. Mix in accordance with the cell suspension: glycerate: 900ul:100ul ratio, -70C stored overnight, stored in liquid nitrogen. When the cells are resuscitation, the cells are quickly thawed in a water bath at 37 degrees C and a suitable amount of complete culture is added. After 5-8 hours, replace the complete culture fluid and conventionally pass on the culture.

Cytotoxicity of IML<br>The lung cancer cells used in this study were routinely cultured in DMEM complete culture medium, which contained 10% fetal bovine serum (FBS) and 1% penicillin streptomycin. The culture conditions were 37 ℃ and 5% CO2. The amount of culture medium: 2 ml for 35 mm, 3 ml for 60 mm and 8 ml for 10 cm.<br>Cryopreservation and resuscitation of cells: the cells were digested with 1.25% trypsin. When the cells became round and had not been floated, a proper amount of complete culture medium was added and repeatedly blown to form cell suspension. Cell suspension: glycerol = 900ul: 100ul was mixed and stored overnight at - 70 ℃ in liquid nitrogen. When the cells were resuscitated, thaw the cells rapidly in 37 ℃ water bath, and then add appropriate amount of complete culture medium. After 5-8 hours, the complete culture medium was changed and passaged routinely.<br>