title: Morphological and ultrastructural evaluation of cultured frozen-thawed human fetal ovarian tissue.
abstract: OBJECTIVE
  To investigate a defined culture condition for the culture of frozen-thawed human ovarian tissue.
  
  
  METHODS
  Prospective laboratory study.
  
  
  METHODS
  Reproductive biology laboratories in university hospitals.
  
  
  METHODS
  Fetal ovarian tissue from elective termination of pregnancy.
  
  
  METHODS
  Culture of frozen-thawed fetal ovarian tissue for up to 63 days.
  
  
  METHODS
  Morphology, morphometry, and survival of follicles in relation to culture times.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  The present culture condition leads to good survival and progressive follicular growth and differentiation that is comparable to the physiological pattern of early folliculogenesis.
authors.affiliation: Follicle Biology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium. jcsadeu@az.vub.ac.be
authors.forename: Jean Clair
authors.initials: JC
authors.lastname: Sadeu
authors.forename: Rita
authors.initials: R
authors.lastname: Cortvrindt
authors.forename: Rafael
authors.initials: R
authors.lastname: Ron-El
authors.forename: Esti
authors.initials: E
authors.lastname: Kasterstein
authors.forename: Johan
authors.initials: J
authors.lastname: Smitz
mesh_terms: D037881:Aborted Fetus / Q000648:ultrastructure*; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D006801:Humans; D010053:Ovary / Q000196:embryology* / Q000648:ultrastructure*; D011247:Pregnancy; D046509:Tissue Culture Techniques