Provided herein are methods of preparing cell therapies under tumor microenvironment (TME) conditions, for example, hypoxic and hyperbaric conditions, for increased cytotoxicity towards cancer cells. Also disclosed herein are improved cell therapies with increased cytotoxicity towards cancer cells. The methods may include producing a CAR-T cell therapy for increased cytotoxicity towards cancer cells, culturing the population of CAR-T cells in the hypoxic environment and the hyperbaric environment for at least 10 generations to produce the CAR-T cell therapy, and the CAR-T cell therapy comprises increased cytotoxicity towards cancer cells as compared to the population of CAR-T cells.

A cryopreserved dairy goat semen metabolite screening and diluent preparation method: preserve semen with base diluent until the day 5, select samples on the days 0, 1, 3 and 5 for metabolome sequencing, screen out gamma-aminobutyric acids decreasing significantly with the preservation time, add different concentrations of gamma-aminobutyric acids to detect sperm viability and motility, and determine the optimal concentration of gamma-aminobutyric acids. The optimal addition concentration of gamma-aminobutyric acids is 1.0 g/L, ensuring that the viability and motility of sperms preserved until the day 5 reach 56.49% and 53.69% and that plasma membrane integrity and acrosome integrity reach 58.93% and 60.12%, and effectively improving the oxidation resistance of sperms and reduce oxidative damage. The invention effectively improves the semen cryopreservation effect, has important significance for reducing breeding stock and improving artificial insemination and reproductive efficiencies in the animal husbandry, high popularization and application prospects and economic values.