Stem cells possess enormous therapeutic potential in tissue replacement. To study stem cells further, they must be isolated. Techniques are available for enrichment and study of hematopoietic stems cells, but thus far, techniques for purification of spermatogonial stem cells have not been described. Enrichment techniques for hematopoietic stem cells include the use of fluorescence-activated cell sorter analysis with Hoechst 33342 and rhodamine 123 (Rho) dyes. Use of Hoechst dye to isolate spermatogonial stem cells has been unsuccessful in our laboratory, and our results have conflicted with those from other laboratories. Taking advantage of the differential staining of the Rho dye, we report a novel method to enrich murine spermatogonial stem cells. Testicular cells are harvested from cryptorchid ROSA26 male mice. Populations of these cells are then stained with the Hoechst and Rho dyes, allowing them to be sorted by flow cytometry into a side population (SP) of Hoechst low-intensity cells and populations of low (Rholow) or high (Rhohi) fluorescent intensity. Sterile recipients, W/Wv mice, with an intrinsic germ cell deficiency were transplanted with the Hoechst SP cells, Rholow, Rhohi, and nonsorted donor cells. No spermatogonial stem cell colonies were derived from the Hoechst SP cells. The number of spermatogonial stem cell colonies from transplanted Rholow cells showed a 17- and 20-fold enrichment over those of Rhohi and nonsorted cells, respectively.
Male infertility presents a public health concern. As most men wish to become fathers, it is important to increase men’s awareness of infertility risk factors. We developed a mobile health application (mHealth app), Infotility XY, to promote men’s reproductive health. This study evaluates whether use of the app led to increased knowledge of infertility risk factors, and whether knowledge change was associated with participants’ sociodemographic characteristics and/or app usage. Participants were recruited between August and October 2020. Eligibility criteria included: identified as male; 18–45 years old; childless; no infertility history; able to read and write in English/French; had internet access. We assessed participants’ fertility knowledge before and after app use. App usage data were captured during the 2-week intervention period. Our sample included 49 men aged 18–45. Seventy-eight percent of participants had not previously sought fertility information. Participants viewed on average 75% of the app’s articles, and 96% of participants said the app increased their fertility knowledge. Before app use, 55% of men said they were aware of infertility risk factors, compared to 96% after app use. Men correctly identified more risk factors after app use compared to before, t(48) = 8.28, p < .001. Participants’ sociodemographic characteristics and amount of app usage were not associated with knowledge change. This study provides evidence of the feasibility of an mHealth app to improve men’s awareness of infertility risk factors. Given the positive relationship between male reproductive health and overall health, increased awareness of infertility risk factors may lead to men’s improved overall health.
While there are a number of studies documenting a positive effect of varicocelectomy on semen parameters, the length of time required following varicocelectomy for semen quality to improve is not well established. Therefore, in this study we identified the changes with time in semen parameters after varicocelectomy.The records and database of 304 patients who underwent varicocele repair for subfertility were reviewed retrospectively. All men had at least 2 preoperative semen analyses as well as semen testing at 3 and 6 months postoperatively.For the 100 patients who met the study inclusion criteria mean sperm counts increased significantly by 3 and 6 months after varicocelectomy (by 53% and 38% by 3 and 6 months, p = 0.0003 and 0.001, respectively). By 3 and 6 months the combined groups had a mean 2.5 and 1.5-fold higher total motile count compared to the preoperative total motile count, respectively. There was no further improvement in semen parameters in those men followed for more than 6 months. There were no statistically significant differences in the improvement in semen volume, motility, count or total motile count comparing the results at 3, 6 and more than 9 months postoperatively.Sperm parameters improve by 3 months after varicocele repair and then do not improve further. This finding should allow physicians to decide quickly if varicocelectomy has been effective and, then, if required, plan on the use of other therapies to manage the couples' infertility.
A committee was established at the request of the Canadian Urological Association to develop guidelines for the investigation and management of azoospermia.Members of the committee, all of whom have special expertise in the investigation and management of male