Experiments for the establishment of drug-resistant cell lines in vivo
Experiments for the establishment of drug-resistant cell lines in vivo
In vivo drug-resistant cell lines are established experimentally by prolonged exposure of cells to low doses of drugs, causing changes in the biochemical processes of the cells themselves and the appearance of Pgp (or P-170) glycoproteins on the cell membranes, which cause the cells to gradually become resistant to the drugs.
Operation method
Drug tolerance test
Principle
Prolonged exposure of cells to low doses of drugs causes changes in the drug chemical processes of the cells themselves, and the appearance of Pgp glycoproteins on the cell membrane, making the cells progressively more tolerant to the drug, which gradually strengthens as the drug concentration is incremented.
Materials and Instruments
Mouse Tumor Strain Move I. Preparation of materials For more product details, please visit Aladdin Scientific website.
Heparin saline
Centrifuge
1. Animal selection: According to the drug-resistant cell line to be established, select suitable animals. During the experiment, animals of the same sex are generally selected for the experiment, and the weight of the animals is selected to be about 17~21 g for females and 18~22 g for males.
2. Selection of tumor strains: common tumor strains are available, and the tumor strains should be determined according to the research purpose and direction. Considering the convenience of generation and easy observation or detection, ascites tumors should be selected.
Experimental Procedures
1. Determination of toxicity of the tested drugs
(1) Randomly divide the mice into 5~8 groups, 10~20 mice in each group, and give each animal a quantitative amount of drug once a day for 5 consecutive days, and then observe for 30 days.
(2) Record the number of deaths of the animals within 30 days. The dose of the drug should be such that the mortality rate of the animals ranges from 0 to 100%.
(3) Calculate the LD10 and the 95% confidence limit that the initial dose at which a resistant strain is established is approximately 1/6 LD10.
2. Tumor cell growth assay
(1) Inoculate each animal with 15×106 cells using ascites cancer cells on day 7 of passaging. 7 days later, execute the animals, open the abdomen, collect all the ascites, and rinse the abdominal cavity three times with saline containing 150 IU of heparin.
(2) Combine the ascites and rinses for tumor cell counting, centrifuge the ascites at 5 500 g for 5 min, and determine the compressed volume of tumor cells.
(3) Tumor cell compression volume was counted in different dosage drug groups and control group, which can measure the effect of drugs on tumor cell growth.
(4) The drug dose is generally at LD10, which can completely inhibit the growth of sensitive strain tumor cells.