Method for culturing primary cancer cells through simulated microgravity-induced reprogramming and applications thereof

A method for simulated microgravity-induced reprogramming of primary cancer cells, comprising:

• obtaining cancerous tissue from a patient with cancer, digesting the cancerous tissue with a digestion solution and washing the cancerous tissue with washing solutions to obtain cancer cells for cultivation, cultivating the cancer cells in a culture medium using a multi-directional G-force generator to simulate a space microgravity environment, wherein the microgravity environment is 10−3 g with a fluctuation range of plus or minus 10%, wherein a cultivation duration in the microgravity environment is 6.5-7.5 days;
• wherein the culture medium used in the method comprises:
• 310-350 ml Dulbecco's Modified Eagle Medium (DMEM), 100-120 ml Ham's F12 nutrient mixture, 40-60 ml fetal bovine serum (FBS), 4-6 ml 200 mM glutamine solution, 4-6 ml penicillin-streptomycin mixture with penicillin at a concentration of 10 kU/ml and streptomycin at a concentration of 10 mg/ml, 400-600 μL hydrocortisone/epidermal growth factor (EGF) solution, 240-260 μL 10 mg/ml insulin, 4-6 μL 25 mg/ml amphotericin B, 90-110 μL 50 mg/ml gentamicin, 0.5-1.5 μL 5 mg/ml cholera toxin, and 480-520 μL 10 mM Y-27632;
• wherein hydrocortisone/EGF solution is prepared by combining 25 μL of 0.1 mg/ml EGF solution with 19 ml of DMEM and 1 ml of 0.5 mg/ml hydrocortisone;
• wherein the cancer is breast cancer;
• wherein the washing solutions used for washing comprise solution A, solution B, solution C, and solution D, wherein the solution A comprises:
• 400-600 ml phosphate-buffered saline (PBS), 4-6 ml penicillin-streptomycin mixture with penicillin at a concentration of 10 kU/ml and streptomycin at a concentration of 10 mg/ml, and 4-6 μl 25 mg/ml amphotericin B;
• the solution B comprises:
• 400-600 ml of the aforementioned culture medium and 8-12 ml penicillin-streptomycin mixture with penicillin at a concentration of 10 kU/ml and streptomycin at a concentration of 10 mg/ml;
• the solution C is penicillin-streptomycin mixture with penicillin at a concentration of 10 kU/ml and streptomycin at a concentration of 10 mg/ml;
• and the solution D is 0.04-0.06 wt % trypsin solution.