Comparative study of testicular sperm maturation in vitro using ham's f10, earle's and ferticult culture media

Testicular sperm extraction [TESE] from azoospermic males followed by intracytoplasmic sperm injection [ICSI] is a recent advance in treatment of male infertility. This study describes the total number and motility changes in vitro of freshly extracted testicular sperm and compares between Ham's F-10, Earle's, and Ferticult culture media. Testicular biopsies were obtained from azoospermic patients [n=11] and processed to obtain a cell suspension that was incubated in equal portions in each culture medium supplemented with 10% human albumin for 5 days. The number of total and motile spermatozoa was evaluated on day 0 and daily afterwards for 5 days. Our results revealed that all patients were suffering from obstructive azoospermia with mean age of 39 +/- 3.36 and duration of infertility of 9.25 +/- 2.98 years. The total number of spermatozoa showed no significant changes in all days of incubation and between all used media [p>0.05]. However, the number of motile spermatozoa was significantly higher in Ferticult medium when compared with Ham's F10 and Earle's media [p <0.05]. In all used media, spermatozoa become motile within 24 hours of culture and the motility was maintained for the 4th day before a decline to the near 0 level is observed on the 5th day of incubation. Furthermore, the number of progressively motile sperm significantly peaked on day 3 of incubation in all media but the best significant yield was observed in Ferticult medium [p <0.05]. No significant correlation was found with age of patients or duration of infertility [p> 0.05]. From our study we recommend that TESE is carried out approximately 3 days before oocyte retrieval and that Ferticult medium is the medium of choice to use in in vitro cultivation for obtaining the most mature of testicular tissue sperm to be used for IVF related procedures, e.g. ICSI

Selective cultivation of epidermal keratinocytes from newborn and adult skin in a serum-free medium and without the use of a feeder layer

Selective cultivation of epidermal human keratinocytes [HK] is important not only in investigating disorders that involve these cells, but also in therapy. In this work we report on a simple modified technique for cultivation of epidermal Hk from newborn and adult skin in which the need for conditioned medium, serum and specialized culture surfaces, e.g., 3T3 cells, has entirely been eliminated. The medium is a modified MCDB 153 to which crude bovine pituitary extract [BPE] 70 ug/ml, epidermal growth factor [EGF] 10 ng/ml, hydrocortisone 1.4 X 10-6 M, and insulin 10 ug/ml were added. The Ca++ level was 0.15 mM, however near confluence the primary culture was passaged and one secondary culture was maintained on this low Ca++ level, while the other was exposed near confluence to high Ca++ level [1.8 mM]. Proliferation rate was calculated in both cultures and each mitogen was omitted from the low Ca + + medium and proliferation rate was also determined at day 3, 6 and 9 of incubation. Cytospin preparation from low and high Ca ++ cultures were labelled with MoAb against cytokeratin 5,14,1 and 10 [APAAP technique]. Colonies were detected after 2 days of inoculation of the primary and secondary cultures and as the cells divided, they expanded and formed a continuous sheet. Cells left without passage underwent morphological changes characteristic of differentiation and were completely separated after 3-4 weeks as a continuous sheet. Cells of secondary cultures exposed to high Ca++ level underwent the same differentiation process and expressed cytokeratins of suprabasal keratinocytes, namely CK1 and 10. On the other hand, cells maintained on low Ca ++ grew as monolayer, were polygonal in shape and expressed cytokeratins similar to basal cells, namely CK 5 and 14

Efficacy of transrectal microwave hyperthermia [TRMH] in treatment of abacterial prostatitis

Thirty patients with chronic abacterial prostatitis were randomly divided into two groups [15 cases each]. Group I was given medical treatment in the form of oral minocycline [100 mg] twice daily for three weeks, whereas group II was given the same medical treatment course combined with eight sessions [1-hour each thrice weekly] of TRMH [43 C] to the prostate. The efficacy of the treatment was evaluated by the effect on symptoms, clinical signs, white blood cells [WBCs] in expressed prostatic fluid [EPF], pre- and post-prostatic massage urinalysis and urodynamic measurements [flowmetry]. The results showed marked improvement of the symptoms and signs among group II patients [excellent results in six, good in three, fair in two and poor in four] and less improvement among group I patients [excellent in three, good in three and poor in nine] as assessed clinically. When a comparison was made between the mean difference of pre- and post-treatment values of groups I and II, superior results were detected with the latter. The mean difference of WBCs in EPF among group II was 19 +/- 9.32 compared with 9.2 +/- 8.2 among group I. Post-prostatic massage urinalysis mean difference in group II was 7.2 +/- 1.99, whereas in group I it was 3.1 +/- 3.93. The uroflowmetric studies showed mean difference Q max to be 4.3 +/- 2.63 in group II, whereas in group I it was 2.0 +/- 1.33. From the above data, it was concluded that transrectal microwave hyperthermia improved the outcome of therapy of chronic abacterial prostatitis and provided a potentially effective therapeutic modality for this condition