This experiment is mainly used to establish in vitro tumor cell models to test drug resistance.

Operation method

Cell Culture Technology

Principle

In vitro drug-resistant tumor cell model establishment experiments take a long time drug treatment, induced resistance, and finally screened out the cell clones with certain resistance.

Materials and Instruments

Mouse
Calf serum DMEM DOX
CO2 incubator Sterile steel ring

Move

I. Cultivation conditions

Cultured with DMEM culture medium containing 10% calf serum, 5% _CO2, 37℃.

Experimental steps

1. CHO cells were cultured in DMEM culture medium containing 10% serum, and DOX (Adriamycin) 0.01 ug/ml was first used to induce the culture of CHO cells for 3 months.

2. Then the concentration of DOX was gradually doubled up to 0.1 ul/ml over 3 months, followed by screening for low concentrations of DOX.

3. Subsequently, the clone _RC1 with higher multiplicity of resistance was ligated with a sterile steel ring to make the dose-response curves of CHO and _RC1 to DOX, and to find the _IC50 values of CHO and _RC1.

4. The multiplicity of resistance is the _IC50 of _RC1 versus the _IC50 of CHO _.

Caveat

1. laboratory conditions for culturing cells should be available to prevent contamination.

2. experimentally calculate the IC of the sensitive cells to the drug.50value to determine the actually induced concentration.

3. Long-term incremental cultures can be used to produce stable drug-resistant cell lines. Cell lines resistant to adriamycin, for example, are often resistant to other naturally occurring antitumor agents.

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