Three-dimensional culture system for screening liver-caner chemotherapeutic drugs
A three-dimensional culture, chemotherapy drug technology, applied in drug screening, tissue cell/virus culture devices, 3D culture, etc., can solve the problem that in vitro cell culture technology cannot perfectly simulate the in vivo environment of tumor tissue, and cannot achieve individualized treatment.
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Embodiment 1
[0024] Embodiment 1, establishment of three-dimensional culture system
[0025] First prepare gel for tissue culture: add 1.5mL coagulant (Adittive 1) containing epidermal growth factor to 12mL matrigel for tissue culture, which contains laminin, collagen IV, fibroblast growth factor , Tissue plasminogen activator, add slowly, mix well while adding, the mixture is light yellow and clear. Add 1.5mL indicator containing phenolphthalein (Adittive 2) into the mixture, add slowly and mix well. When just added, it can be seen that the local solution is pink, and after mixing, it becomes light yellow and clear. After mixing the components, it is prepared into 15mL tissue culture gel.
[0026] Add the prepared gel into a 48-well culture plate, and add 250uL of gel to each well. Place it in a clean bench and wait for the liquid gel to become translucent.
[0027] Extract surgically resected liver cancer tissue samples, place the tissue in PBS solution, and separate into 0.5cm 3 siz...
Embodiment 2
[0030] Example 2. Using a three-dimensional culture system to detect the sensitivity of liver cancer tissue to chemotherapeutic drugs
[0031] Prepare the chemotherapeutic drug to be tested by using culture medium or PBS, and keep the ratio of the volume of the medium to the volume of the chemotherapeutic drug greater than 100 according to the concentration indicated in the drug instruction.
[0032] The tumor tissue pieces were cultured in the culture system for 24h, 48h, 72h, 96h and 120h, and the cell growth was observed every day, and the cell migration distance was measured.
[0033] Collect relevant data. Statistical analysis of the effects of different drugs on tumor cell growth and migration.
[0034] We used the above-mentioned three-dimensional tissue culture technology to study the sensitivity of hepatocellular carcinoma tissues to chemotherapy drugs ( Figures 2A-2F ).
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