No abstract

Please cite this paper as: Balen A, Harris S, Chambers E, Picton H. Conservation of fertility and oocyte genetics in a young woman with mosaic Turner syndrome. BJOG 2010;117:238-242. Case reportWe report a case of oocyte cryopreservation and ovarian tissue cryopreservation in a woman with mosaic Turner syndrome in whom, for the first time in the literature, a detailed assessment was made of the genetic composition of the oocytes. The aim was to preserve fertility in a woman considered at high risk of premature ovarian failure.The patient presented at the age of 12 years with short stature. Her parents had been concerned about her rate of growth compared with her contemporaries. She had no other stigmata of Turner syndrome, but investigations confirmed mosaicism with 45,X/46,XX, with only two of the 30 cells examined being 46,XX and the remainder being 45,X. All other investigations were normal and, in particular, an ultrasound scan revealed a prepubertal uterus with ovaries clearly visualised and of normal volume.Treatment was commenced with growth hormone under the care of a paediatric endocrinologist. This produced a good response with a final height of 1.55 m. Puberty occurred naturally and menarche was achieved at the age of 15 years and 0 months, followed by a regular menstrual cycle. There was fluctuating thyroid function and, eventually, the girl became hypothyroid and is now on thyroxine replacement.The patient consulted at the age of 17 years to talk about the possibility of ovarian cryopreservation. At that stage, we were undertaking research into ovarian tissue cryopreservation for women about to undergo chemotherapy. 1,2 Therefore, counselling was provided with regard to the risks and limitations of this process. 3 The patient continued to have a regular menstrual cycle with normal gonadotrophin concentrations and, therefore, in February 1998, laparoscopy was performed and bilateral ovarian biopsies were taken. Both ovaries appeared to be of normal size and appearance. Multiple samples of ovarian cortical tissue were collected laparoscopically using punch biopsy and the tissue was cryopreserved by slow freezing following equilibration in 1.5 m dimethylsulphoxide and 0.5 m sucrose, according to the protocol described by Newton et al., 1 and stored under liquid nitrogen. A specimen from each ovary was karyotyped. The analysis of cultures from the left ovarian biopsy showed 16 cells with a normal female pattern and 14 cells with only a single X chromosome. From the right ovary, eight cells had a normal female pattern and 22 cells had a single X chromosome. Therefore, these ovarian tissue biopsies showed a significantly higher proportion of cells with a normal female pattern. The patient was reviewed annually and continued to have a fairly regular 28-30-day menstrual cycle with normal serum gonadotrophin concentrations.At the age of 25 years, the serum follicle-stimulating hormone concentration was 3.3 iu/l with a luteinising hormone level of 1.3 iu/l. A measurement of inhibin B yielded 176 pg/ml. These ...

30 October 2009.

Serum AMH has the capacity to act as a diagnostic test for PCOS. Moreover, since its value rises with the more marked phenotypes, different cut-off values need to be used to differentiate those patients with polycystic ovarian morphology (PCOM), hyperandrogenism (HA) and oligoanovulation (OA).

The regulation of pig theca cell steroidogenesis was studied by the development of a physiological serum-free culture system, which was subsequently extended to investigate potential theca-granulosa cell interactions. Theca cells were isolated from antral follicles 6-9 mm in diameter and the effects of plating density (50-150x10(3) viable cells per well), LH (0.01-1.0 ng ml(-1)), Long R3 insulin-like growth factor I (IGF-I) (10, 100 ng ml(-1)) and insulin (1, 10 ng ml(-1)) on the number of cells and steroidogenesis were examined. The purity of the theca cell preparation was verified biochemically and histologically. Co-cultures contained 50x10(3) viable cells per well in granulosa to theca cell ratio of 4:1. Wells containing granulosa cells only were supplemented with 'physiological' doses of androstenedione or 100 ng ml(-1). Oestradiol production by co-cultures was compared with the sum of the oestradiol synthesized by granulosa and theca cells cultured separately. Oestradiol and androstenedione production continued throughout culture. High plating density decreased steroid production (P < 0.01). LH increased androstenedione (P < 0.001) and oestradiol (P < 0.05) synthesis and the sensitivity of the cells increased with time in culture. Oestradiol production was increased by 10 ng IGF-I ml(-1) (P < 0.001) but androstenedione required 100 ng ml(-1) (P < 0.001). Co-cultures produced more oestradiol than the sum of oestradiol synthesized by theca and granulosa cells cultured separately (P < 0. 001), irrespective of the androstenedione dose. This serum-free culture system for pig theca cells maintained in vivo steroidogenesis and gonadotrophin responsiveness. Thecal androstenedione and oestradiol production were differentially regulated and were primarily stimulated by LH and IGF-I, respectively. Theca-granulosa cell interactions stimulated oestradiol synthesis and this interaction was mediated by factors additional to the provision of thecal androgen substrate to granulosa cells.