194 Carl A. Gemzell 



SUBJECTS AND METHODS 



Ovulation was induced in young women with primary or secondary 

 amenorrhea with an FSH preparation obtained from human pituitaries 

 and with a Swedish commercial preparation of human chorionic gonado- 

 tropins (Gonadex-Leo). All the patients were treated with human chorionic 

 gonadotropin (HCG) several months before the administration of FSH in 

 order to exclude the possibility that HCG alone could induce ovulation. 



Preparation of human pituitary FSH. Human pituitaries obtained from 

 autopsy cases were frozen and lyophilized. The dried glands were cut into 

 small pieces and extracted in cold CaO-solution at pH 9.3 under continuous 

 stirring. After centrifugation the clear supernatant was brought to 55% 

 saturation by the addition of saturated ammonium sulfate. The precipitate 

 was discarded and the clear supernatant was brought to 75% saturation by 

 the addition of solid ammonium sulfate. The precipitate was collected by 

 centrifugation, dissolved in water, dialyzed and lyophilized. This product 

 was called human pituitary FSH and was used in the clinical trials. 



Potency of human pituitary FSH. The human pituitary FSH was assayed 

 against the provisional human menopausal gonadotropin (HMG-20A) 

 standard preparation. On a weight basis the partially purified human pituitary 

 FSH was thirty to fifty times as potent as the HMG-20A standard preparation 

 when assayed by methods measuring total gonadotropin or FSH activities. 

 In the ventral prostate test, which is considered to be specific for LH activity, 

 the human pituitary FSH was only approximately 5 times as potent as 

 HMG-20A. 



Purification of human pituitary FSH. A further purification of the human 

 pituitary FSH was carried out by Dr. P. Roos in Uppsala employing ion 

 exchange chromatography and zone electrophoresis. A fraction was obtained 

 which is more than 2000 times as active as HMG-20A. 



Achninistration of human pituitary FSH. The human FSH preparation was 

 administered in daily doses of 10 mg during a 10-day period. The 10 mg dose 

 was chosen as it gave a significant increase in ovarian size and estrogen 

 excretion in a hypopituitary dwarf. A dose of 1 mg per day had no effect 

 and doses of 2 and 5 mg gave only a slight increase in proliferative endometrial 

 activity. The 5 mg dose was tried in several cases without any effect. It 

 may be suggested that a relatively large dose has to be administered in 

 order to initiate the follicular growth; later a smaller dose may be enough. 

 Thus, the effective daily dose of FSH corresponded to about 500 mg of the 

 HMG-20A standard and the total dose during the 10-day period to about 

 5000 mg. 



Repeated treatments with human pituitary FSH were performed in at 

 least 10 women. All of these 10 women responded to the first treatment as 

 well as to the subsequent ones. A young woman with primary amenorrhea 

 was treated during a period of 2 years and received more than 1 gm of FSH. 



