K Oktay1, H Newton, R G Gosden. 1. Cornell University-Weill Medical College and New York Methodist Hospital, New York, New York 11215, USA.
Abstract
OBJECTIVE: To determine the long-term survival of frozen-thawed human ovarian tissue as xenografts in severe-combined-immunodeficiency (SCID) mice. DESIGN: Animal study. SETTING: Animal and laboratory facilities at an academic center. PATIENT(S): Ovarian tissue obtained from a 27-year-old woman. INTERVENTION(S): Grafting of frozen-thawed ovarian tissue in SCID mice for 22 weeks. MAIN OUTCOME MEASURE(S): Follicle counts and growth by morphology and PCNA staining in frozen-thawed grafts and fresh controls. RESULT(S): All three grafts were recovered intact after 22 weeks. Their stroma was devoid of necrotic cells and contained healthy follicles. The ratio of primordial-total follicles decreased significantly after grafting (0.94 +/- 0.02 to 0.87 +/- 0. 01, control vs. grafting). Compared with controls, after 22 weeks of grafting, a higher percentage of follicles had initiated growth (5.6 +/- 2.4 vs. 12.5 +/- 1.9), but there was still a significant number of primordial follicles/graft (75 +/- 6.8). Follicle stages were similar between two groups; only primordial and one-layer follicles were seen in the xenografts. In the controls, except for one two-layer follicle, the most advanced follicle was at the one-layer stage. CONCLUSION(S): Human primordial follicles survive freeze-thaw and long-term xenografting procedures and retain their capacity to initiate growth. These findings encourage future attempts for human autologous ovarian transplantation.
OBJECTIVE: To determine the long-term survival of frozen-thawed human ovarian tissue as xenografts in severe-combined-immunodeficiency (SCID) mice. DESIGN: Animal study. SETTING: Animal and laboratory facilities at an academic center. PATIENT(S): Ovarian tissue obtained from a 27-year-old woman. INTERVENTION(S): Grafting of frozen-thawed ovarian tissue in SCIDmice for 22 weeks. MAIN OUTCOME MEASURE(S): Follicle counts and growth by morphology and PCNA staining in frozen-thawed grafts and fresh controls. RESULT(S): All three grafts were recovered intact after 22 weeks. Their stroma was devoid of necrotic cells and contained healthy follicles. The ratio of primordial-total follicles decreased significantly after grafting (0.94 +/- 0.02 to 0.87 +/- 0. 01, control vs. grafting). Compared with controls, after 22 weeks of grafting, a higher percentage of follicles had initiated growth (5.6 +/- 2.4 vs. 12.5 +/- 1.9), but there was still a significant number of primordial follicles/graft (75 +/- 6.8). Follicle stages were similar between two groups; only primordial and one-layer follicles were seen in the xenografts. In the controls, except for one two-layer follicle, the most advanced follicle was at the one-layer stage. CONCLUSION(S): Human primordial follicles survive freeze-thaw and long-term xenografting procedures and retain their capacity to initiate growth. These findings encourage future attempts for human autologous ovarian transplantation.