Human Reproduction, Vol 12, 1032-1036, Copyright © 1997 by Oxford University Press
O Hovatta, R Silye, R Abir, T Krausz and RM Winston
Ovarian cortical tissue was obtained during gynaecological operations by
biopsy or after oophorectomy from 20 women aged 25-42 years. It was placed
in organ culture, either fresh or following thawing after cryopreservation,
for 1-4 months. The tissue was cut in slices 0.1-0.3 mm in diameter and
transferred to 12 mm inserts in 24-well culture plates. These slices were
cultured for 4-21 days in either alpha minimum essential medium (alpha-MEM)
or Earle's balanced salt solution with added pyruvate. Both media were
supplemented with 10% human serum, insulin, gonadotrophins and antibiotics.
Half of the inserts were precoated with extracellular matrix (Matrigel).
Histological samples revealed that there were viable, non-atretic,
primordial, primary and secondary follicles in all the cultures. Mitoses
were seen in the granulosa cells of the secondary follicles. Although the
proportion of atretic follicles increased during culture, non-atretic
follicles were still present after 21 days. After 4-11 days the proportion
of viable follicles was significantly higher when cultured in Earle's
solution supplemented with pyruvate, than when cultured in MEM (77 versus
38%, P < 0.001). In cultures with extracellular matrix the proportion of
viable follicles was significantly higher after 10-15 days than it was
without matrix (85 versus 19%, P < 0.001). Culture after thawing frozen
ovarian tissue did not affect the density or the proportion of the viable
follicles. Two-thirds of follicles in cryopreserved tissue were viable
after 10-15 days in culture. The results indicate that it is possible to
culture human primary and primordial follicles in vitro, and follicles in
cryopreserved tissue are viable.
ARTICLES
Extracellular matrix improves survival of both stored and fresh human primordial and primary ovarian follicles in long-term culture
Infertility Clinic, The Family Federation of Finland, Helsinki.
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