Human Fetal Follicles Research Articles

Tissue culture before transplantation of frozen-thawed human fetal ovarian tissue into immunodeficient mice

To evaluate the effects of tissue culture on the viability and development of follicles in frozen-thawed human fetal ovarian tissue before transplantation into severe combined immunodeficient (SCID) mice and to determine the optimal duration of pretransplant tissue culture. Experimental prospective study. Animal center and reproductive laboratories in university hospitals. Frozen-thawed human fetal ovarian tissue samples from 20-week-old abortuses were randomly divided into four groups and cultured in vitro for 0, 3, 6, or 9 days before being xenografted into kidney capsules of bilaterally oophorectomized severe combined immunodeficient (SCID) mice. Grafts were removed 16 weeks after transplantation. Histological analysis and assessment of proliferative cell nuclear antigen (PCNA) expression levels were used to evaluate the survival and development of follicles. The proportion of growing follicles was significantly increased in groups cultured before transplantation as compared with the noncultured group. Sixteen weeks after transplantation, the number of follicles in the cultured grafts was higher than that in the noncultured grafts. Grafts cultured for 6 or 9 days showed higher proportions of preantral and antral follicles than grafts cultured for 0 or 3 days. PCNA immunohistochemical analysis indicated that follicle cells were in a proliferative state after culture and transplantation. The viability and development of human fetal follicles may be improved by pretransplant tissue culture. The optimal culture duration before transplantation of fetal ovarian tissue is 6 days.

Transplantation of frozen–thawed late-gestational-age human fetal ovaries into immunodeficient mice

To compare the development of human fetal follicles from late-gestational-age fetuses frozen-thawed gradually and slowly with dimethylsulfoxide (DMSO) or 1,2 propanediol (PROH) and sucrose after renal grafting into follicle stimulating hormone-treated immunodeficient mice. Controlled histologic study of grafted human fetal ovaries. Major tertiary care academic center. Eleven women undergoing pregnancy termination at 22 to 33 gestational weeks. None. Microscopic morphometric analysis and immunohistochemistry for proliferating cell nuclear antigen (PCNA). Only follicles from samples frozen-thawed with PROH developed to secondary and antral stages 4 to 6 months after grafting, with PCNA expression in their granulosa cells. However, the number of surviving/developing follicles per section was very low (4-25 per graft), compared with 71 to 406 follicles in pretransplantation samples. Graft recovery was very high, with similar rates for transplants frozen-thawed with PROH and DMSO. Normal ovarian structure after grafting was identified only in the PROH frozen-thawed grafts. In deteriorated grafts, frozen-thawed with either DMSO or PROH, net-like hollows replaced follicles, whereas tubule-like structures were only identified in DMSO frozen-thawed grafts. This is the first report of the development of late-pregnancy-stage human fetal follicles in immunodeficient mice. PROH freezing-thawing supported development and survival better than DMSO. However, the low follicular survival points to the urgent need for efficient methods to enhance vascularization rate and prevent ischemia.

Parameters affecting successful transplantation of frozen-thawed human fetal ovaries into immunodeficient mice

Objective To compare the development and survival of human fetal follicles frozen-thawed with dimethylsulfoxide (DMSO) and propandiol (PROH) in immunodeficient mice, to study the effects of host treatment with FSH, and to compare kidney and subcutaneous transplantation. Design Controlled histologic study. Setting Major tertiary care and referral academic center. Patient(s) Twenty-one women undergoing second-trimester pregnancy termination. Main outcome measure(s) Microscopic morphometric analysis and immunocytochemistry for proliferating-cell nuclear antigen in human fetal ovaries grafted into immunodeficient mice. Result(s) Renal grafts that were frozen-thawed with DMSO rather than PROH survived better in the hosts (79.6% compared with 58.8%), but significantly more follicles were identified in grafts frozen-thawed with PROH ( P<.001). Follicular development was observed only in FSH-treated hosts, and follicular survival and development was better in the kidney than the subcutaneous site. Conclusion(s) This is the first report showing development of human fetal follicles in immunodeficient mice. Freezing-thawing with PROH seems to support development and survival better than with DMSO. The kidney is a better transplantation site than the subcutaneous site, probably because of its superior vascularization. Administration of FSH to the host is essential for follicular development. Follicular development and growth was better in ovarian grafts from older fetuses, as they contained more formed follicles.

Development of human fetal follicles in an immunodeficient mouse.

Development of human fetal follicles in an immunodeficient mouse.