RESUMO
Objective: To describe development of a methodology for an outcome study of children born following in-vitro fertilization or spontaneously-conceived, as a model for defining normal and below-normal development of school-age children for research purposes. Study Design: The main issues addressed were defining the major health and developmental domains to be investigated, selection of age-appropriate validated instruments, considering time constraints to maximize compliance, and budgetary limitations. The final protocol included a half-hour structured telephone interview with mothers of all 759 children and a 2-h developmental assessment of 294 of them. Each of the instruments and recruiting methods are described in terms of the abovementioned considerations. Results: Almost all of the mothers who agreed to be interviewed completed it within the half-hour allotted; however only about half of those who agreed to bring the child for the developmental assessment actually did so. The entire examination battery, assessing cognitive ability, executive functions, attention, and learning skills, was completed by almost all 294 children. There was a significant degree of agreement between the maternal report of the child's reading, writing and arithmetic skills and the in-person examination, as well as regarding the child's weight and height measurements. Conclusion: The findings lend support for a low-budget study, relying on telephone interviews. However, limitations such as the validity of maternal report and recall bias must be taken into consideration.
RESUMO
OBJECTIVE: To investigate a defined culture condition for the culture of frozen-thawed human ovarian tissue. DESIGN: Prospective laboratory study. SETTING: Reproductive biology laboratories in university hospitals. PATIENT(S): Fetal ovarian tissue from elective termination of pregnancy. INTERVENTION(S): Culture of frozen-thawed fetal ovarian tissue for up to 63 days. MAIN OUTCOME MEASURE(S): Morphology, morphometry, and survival of follicles in relation to culture times. RESULT(S): The proportion of primordial, early primary, and primary follicles in frozen-thawed (day 0) ovarian tissue was 77.5%, 21.7%, and 0.8%, respectively. Pronounced degeneration was found in all cell types, and < or =36% of the follicles had signs of atresia at days 7-14, but this figure improved with culture time to <20% of the total follicular population. After 7-14 and 21-35 days of culture, the relative proportion of the follicles in the different classes remained nearly stable. Morphometric examination of healthy follicles showed a significant increase in both follicle and oocyte diameter compared with control. A few follicles had developed to the early secondary stage. Ultrastructural analysis demonstrated well-preserved morphological integrity of healthy primordial and early primary follicles. Immunohistochemical localization of proliferating cell nuclear antigen was positive in proliferating follicular cells at days 7-14 and 21-35 of culture. CONCLUSION(S): The present culture condition leads to good survival and progressive follicular growth and differentiation that is comparable to the physiological pattern of early folliculogenesis.
