Biomedical subjects
Publications and source records attributed to H Picton.
Despite early success with sperm and embryo cryopreservation, human oocyte cryopreservation has met with limited success both in terms of survival and subsequent fertilization. However, compared with embryo cryopreservation, oocyte freezing has great potential for eliminating the ethical and legal problems of embryo storage.
As survival rates for young cancer patients continue to improve, protection against iatrogenic infertility caused by chemotherapy and/or radiotherapy assumes a higher priority. As things stand, women patients have few options to preserve their fertility while children have none at all.
A culture system has been designed in which enzymatically isolated oocyte-granulosa cell complexes from fresh and frozen-thawed ovine ovarian tissue can be grown to antral size in vitro. Oocyte-granulosa complexes ranging from 100 to 240 microns in diameter were dissected from stromal tissue and grown individually in serum-free medium for 30 days. Complexes < 190 microns generally excluded their oocytes or lost three-dimensional structure early in the culture period. In contrast, complexes isolated from fresh or frozen-thawed tissue and measuring 190-240 microns on the day of isolation formed antral cavities in 25 +/- 9% and 18 +/- 6% (mean +/- SEM) of cases, respectively. The effect of gonadotrophin supplementation to the culture medium was tested on frozen-thawed oocyte-granulosa cell complexes only. In cultures supplemented with both FSH and LH or FSH alone, there was no significant difference in the number of oocyte-granulosa cell complexes that formed antral cavities (18 +/- 7%). However, antrum formation was significantly less frequent in cultures lacking gonadotrophin stimulation (7 +/- 4%). All oocyte-granulosa cell complexes maintained a three-dimensional structure throughout culture and developed a functional P450 aromatase enzyme complex, as revealed by the induction of oestradiol production during 8 days of culture after antrum formation in serum-free medium containing testosterone. Oocytes recovered after 30 days of culture were viable and had increased in diameter from 78 +/- 2 microns on the day of isolation, to 131 +/- 3 microns at the end of culture. These results show that oocyte-granulosa cell complexes isolated from cryopreserved ovarian tissue can be grown to antral size in vitro with similar efficiency to those isolated from fresh tissue.
The mammalian oocyte is formed early in foetal life and may remain suspended at diplotene of the first meiotic prophase until several decades later. When, following follicular recruitment, the oocyte enters a growth phase where it increases in volume, and undergoes replication and redistribution of the cytoplasmic organelles. These modifications, including secretion of the zona pellucida, reflect a period of intensive RNA synthesis as the oocyte grows and accumulates the molecular program for embryogenesis. Throughout oocyte development follicle cell support is fundamental to provide the germ line cell with nutrients and growth regulators to ensure progression through the protracted growth phase. Conversely, the oocyte actively promotes growth and differentiation of the follicular cells. Finally, re-initiation of meiosis in mature oocytes results in the production of haploid gametes which are capable of supporting early embryo development. Our present knowledge of the molecular biology of mammalian oogenesis is discussed in this review.