Pairs of fragmented ovarian tissue (with or without MHY1485 treatment) from the same donor were cultured for 2 days in vitro with a medium change every 24 h. Then, the host animals were anesthetized and fragments were randomly transplanted under separate sides of the kidney capsule in the same ovariectomized adult (6–8-week-old) SCID mice. Three days after transplantation, hosts received daily i.p. injections of FSH (1 IU/day) for 28 days to promote follicle development. Thirty days after transplantation, grafts from the same recipient were recovered and weighed before histological analyses. Furthermore, we transplanted the activated fragments into ovariectomized adult SCID mice so that we could dynamically monitor the hormone levels.

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