Effects of in vitro storage temperature and duration time on the state and function of primary granulosa cells after isolated from human follicular fluid

Abstract From the collection of human follicular fluid to the isolation of primary granulosa cells, the duration of these two time points is variable. And there is no specific experiment to explore which temperature will make the cell state better in subsequent experiments. In this study, the collected follicular fluid was divided into 7 groups: one fresh control group, three in vitro storage temperatures (4℃, Room Temperature, and 37℃) and two storage duration times (4h and 8h). After isolating the cells, they were cultured in vitro to evaluate their state and function. The results showed that granulosa cells of the fresh control group had better cell viability and steroid hormone secretion ability than the other treatment groups; with the prolongation of in vitro storage time, the cell viability and steroid hormone secretion ability have decreased to varying degrees; low-temperature in vitro storage is conducive to the maintenance of cell survival and steroid hormone secretion ability, which is also confirmed by the detection of ROS, which can reflect the oxidative stress state of cells. This study showed that after obtaining follicular fluid, the in vitro storage time should be shortened as much as possible, and stored at low temperature, which can keep cells maintain good activity and steroid hormone secretion ability, and reduce oxidative stress and aging. It is of great significance for improving the state of human primary granulosa cells after isolation and subsequent experiment, and it also has certain guiding significance for standardization experimental research programs.