1978 Volume 69 Issue 12 Pages 1590-1604
The effect of glucose, testosterone and FSH on spermatogenesis was studied in organ cultures of 21-day-old rat testes by modification of Trowell's method. The control medium used in all cultures consisted of MEM (Eagles minimum essential medium), supplemented with 10% fetal calf serum, 1mM of sodium pyruvate, 0.1mM each of alanine, aspartic acid, asparagine, glutamic acid, glycine, proline and serine, and 3mM of glutamine and penicillin 100U, per ml. The culture medium was maintained at PH7.1 and explants were incubated at 31°C in a gas atmosphere of 5% CO2 and 95% air.
The results are summarized as follows.
1) In the control medium, primary spermatocytes showed a marked dimution in number during 4 to 7 days in culture, and gradually disappered in almost all tubules within 14 days in culture, even though spermatogonia were well maintained and mitotic activity continued after 42 days in culture.
2) Sertoli cells in all tubules were well maintained at an almost definite number of 16 to 19 per seminiferous tubules for as long as 42 days.
3) Effects of glucose:
In cultures grown in 2mg/ml concentration of glucose medium, the number of primary spermatocytes was slightly greater than in the control medium containing 1mg/ml of glucose for 7 days in culture. In cultures grown in a 3mg/ml concentration of glucose medium, however, the seminiferous tubules showed moderate diminution in diameter and moderate thickening of the tubular wall with the number of primary spermatocytes being smaller than in the control medium.
4) The osmotic pressure in the culture medium was evaluated to study the toxic effect of a higher concentration of glucose on the maintenance of primary spermtocytes. The osmotic pressure being 330mosm/L in the 2mg/ml concentration of glucose medium and 335mosm/L in the 3mg/ml concentration, the osmotic pressure was proved to be within 330mosm/L in view of the maintenance of primary spermatocytes and the structure of seminiferous tubules.
5) Effect of testosterone:
The concentration of testosterone in culture medium for the best maintenance of primary spermatocytes was 50μg/ml concentration, and 5μg/ml, 100μg/ml, in that order.
6) Effect of NIAMDD-rat FSH, B-1:
Addition of FSH to the control medium resulted in good maintenance in the number of primary spermatocytes. A 200μg/ml concentration of FSH was more effective for the maintenace of primary spermatocytes than 100μg/ml of FSH.
As mentioned above, the most effective culture medium for the maintenance of primary spermatocytes was proved to be 2mg/ml of glucose, 50μg/ml of testosterone and 200μg/ml of FSH.
