The Effect of Human Follicular Fluid on Embryonal Development of Mouse in In Vitro Culture / 대한불임학회지

The follicular fluid (FF) of ovary contains various biological active products which affected on the growth of follicles and the fertilization of oocyte in physiological reproductive process of mammals. This study was designed to determine the effects of human FF on fertilization of oocyte and embryonal development in vitro culture. The FF was prepared as clear without blood contamination by needle aspiration from mature follicles of human at the time of oocytes retrieval for in vitro fertilization (IVF). As the medium for culture in vitro of embryonal cells, human tubal fluid (HTF) supplemented with follicular fluids at concentrations of 10%, 40% and pure FF were used. These effects were compared to control group of cultured embryos in HTF supplemented with 0.4% BSA (bovine serum albumin). For IVF, 64 eggs in control group, 67 eggs in 10% FF, 57 eggs in 40% FF and 64 eggs in pure FF were respectively allocated. And the rates of fertilization were almost similar in all groups as resulting 82.81% in control, 85.07% in 10% FF, 87.71% in 40% FF and 81.25% in pure FF. On the examination for embryonal cleavage from fertilized eggs, the rates of developing to 4 cell stage was similar in all groups, as results 98.11% in control, 98.27% in 10% FF and 98% in 40% FF but 78.84% in pure FF. And the rates of developing to 8-16 cell stage were significantly reduced as 44% in 40% FF and 44.23% in pure FF (p<0.05) compare to 71.69% in control media. As likewise, the rates of developing to morular stage were also significantly reduced to 36% (p<0.05) and 21.15% (p<0.01) respectively in 40% FF and pure FF And the rates to blastocystic stage of embryo was lowest as 7.69% in pure FF. The quality of embryonal cells on cleavage to the 8-16 cell stage was poorer, higher concentrations of FF The rates of grade 1 in pure FF, as 23.07%, was lowest compare to those of other groups, in which the rates of grade 1 in control, 10% FF and 40% FF were 58.49%, 47.36% and 34% respectively. And on the contrary, the rate of grade 4 in pure FF was highest as 23.07%, while those were 5.66% control, 8.77% in 10% FF and 20% in 40% FF. On the viability of embryos, the rate of embryonal cell death was more rise, at the higher concentrations as well as longer exposure in the follicular fluid. At 48 hours after in vitro culture of embryos, the rate of survival embryos in pure FF was markedly lowered as 44.23%, compare to that of control (p<0.05). But there was not significant difference between the rates of survival embryos in each group beside the pure FF, which the rates were 77.35% in control, 70.17% in 10% FF and 60% in 40% FF respectively. And at 72 hours after in vitro culture, the rates of survival embryos were also significantly dropped to 21.15% in pure and 36% in 40% at concentration of FF compare to 62.26% in control (p<0.05, p<0.01). Finally, the rate of embryonal death at 96 hours after in vitro culture was highest as 82.69% in pure FF among all groups which those were 35.84 in control, 56.14% in 10% FF and 64% in 40% FF respectively. In conclusion, this study suggests that the FF has no effects, in particular, to the in vitro fertilization of oocytes but exerted a bad effect to the cleavage, quality and viability of the embryonal cells during in vitro culture. However, the FF is harmful on embryonal development at conditions in higher concentration and especially on the embryos after 8~16 cell stage.

Double Tracheoesophageal Fistula

Esophageal atresia(EA) with a double tracheoesophageal fistula(TEF) is rare. It accounts for only 0.7% of all cases of EA and TEF. A male newborn weighing 2860g was born by normal vaginal delivery at 41weeks' gestation to a 27-year-old mother who had a normal pregnancy. But immediately developed recurrent choking and respiratory distress. Feeding tube was inserted and chest X ray showed the feeding tube coiled in the proximal pouch(T-2 level). With a preoperative diagnosis of EA and distal TEF, the thoracotomy was performed on the third day of life. At thoracotomy, TEF was proved to be a double fistula. Both fistulas were divided and an esophageal anastomosis was performed. The postoperative course was uncomplicated until the eighth postoperative day when a minor anastomotic leak. The patient was discharged at 103 days of age.

Bilateral Adrenal Neuroblastoma

Neuroblastoma is the most common solid malignancy in childhood. However, a neuroblastoma presenting as bilateral and multifocal is very rare. A male newborn weighing 2.7 kg was born by normal vaginal delivery at 39 weeks' gestation to a 27-year-old mother who had a normal pregnancy. He was in good condition at birth but presented palpable masses in the both upper abdomen and both side of the neck. Ultrasound examination showed the lesions to be 3 × 3cm sized calcified mass in right suprarenal area and 5 × 3cm sized homogenous mass in left suprarenal area. The abdominal mass was also examined by computed tomography with similar findings. With the impression of bilateral neuroblastoma or metastatic spread, the laparotomy was performed on the 13th day of life. Frozen section of biopsy of the left neck mass was obtained, which showed neuroblastoma. Intraoperative findings revealed bilateral adrenal masses which were distinct and anatomically separate. Bilateral adrenalectomy was accomplished and the initial postoperative course was unevenful. The patient was discharged for hopeless and expired at age 45 days. We present bilateral adrenal neuroblastoma considered to be simultaneous occurrence rather than metastases from one site to another.