Tissue Collection
- For this investigation, ovaries were collected from a local Greeley slaughterhouse and transferred to the lab in 1X PBS
- The corpus luteum was carefully dissected from the ovaries and cut into approximately 1 mm slices
- These slices were kept in Hank’s balanced salt solution (HBSS)
- The pieces were then moved to culture flasks with bovine serum albumin (BSA) and type one collagenase. These flasks were then incubated in a moving water bath at 37 ℃ for one and a half hours
- The samples were filtered to remove debris such as connective tissue and spun at 500 g. The remaining tissue was transferred to T-75 flasks and resuspended in Hams F-12 media with FBS, insulin transferrin selenium, and antiviral-antibacterial supplements