Maintenance of hPSCs under Xeno-Free and Chemically Defined Culture Conditions

Previously, the majority of human embryonic stem cells and human induced pluripotent stem cells have been derived on feeder layers and chemically undefined medium. Those media components related to feeder cells, or animal products, often greatly affect the consistency of the cell culture. There are clear advantages of a defined, xeno-free, and feeder-free culture system for human pluripotent stem cells (hPSCs) cultures, since consistency in the formulations prevents lot-to-lot variability. Eliminating all non-human components reduces health risks for downstream applications, and those environments reduce potential immunological reactions from stem cells. Therefore, development of feeder-free hPSCs culture systems has been an important focus of hPSCs research. Recently, researchers have established a variety of culture systems in a defined combination, xeno-free matrix and medium that supports the growth and differentiation of hPSCs. Here we described detailed hPSCs culture methods under feeder-free and chemically defined conditions using vitronetin and TeSR-E8 medium including supplement bioactive lysophospholipid for promoting hPSCs proliferation and maintaining stemness.

Usage and perceptions of phosphodiesterase type 5 inhibitors among the male partners of infertile couples / 대한생식의학회지

OBJECTIVE: We aimed to investigate the prevalence of erectile dysfunction (ED) and the usage of phosphodiesterase type 5 (PDE5) inhibitors for ED treatment in infertile couples. METHODS: A total of 260 male partners in couples reporting infertility lasting at least 1 year were included in this study. In addition to an evaluation of infertility, all participants completed the International Index of Erectile Function (IIEF)-5 questionnaire to evaluate their sexual function. The participants were asked about their use of PDE5 inhibitors while trying to conceive during their partner's ovulatory period and about their concerns regarding the risks of PDE5 inhibitor use to any eventual pregnancy and/or the fetus. RESULTS: Based on the IIEF-5 questionnaire, 41.5% of the participants (108/260) were classified as having mild ED (an IIEF-5 score of 17-21), while 10.4% of the participants (27/260) had greater than mild ED (an IIEF-5 score of 16 or less). The majority (74.2%, 193/260) of male partners of infertile couples had a negative perception of the safety of using a PDE5 inhibitor while trying to conceive. Only 11.1% of men (15/135) with ED in infertile couples had used a PDE5 inhibitor when attempting conception. CONCLUSION: ED was found to be common in the male partners of infertile couples, but the use of PDE5 inhibitors among these men was found to be very low. The majority of male partners were concerned about the risks of using PDE5 inhibitors when attempting to conceive. Appropriate counseling about this topic and treatment when necessary would likely be beneficial to infertile couples in which the male partner has ED.

Reversible Infertility Associated with Testosterone Therapy for Symptomatic Hypogonadism in Infertile Couple

PURPOSE: Androgen replacement therapy has been shown to be safe and effective for most patients with testosterone deficiency. Male partners of infertile couples often report significantly poorer sexual activity and complain androgen deficiency symptoms. We report herein an adverse effect on fertility caused by misusage of androgen replacement therapy in infertile men with hypogonadal symptoms. MATERIALS AND METHODS: The study population consisted of 8 male patients referred from a local clinic for azoospermia or severe oligozoospermia between January 2008 and July 2011. After detailed evaluation at our andrology clinic, all patients were diagnosed with iatrogenic hypogonadism associated with external androgen replacement. We evaluated changes in semen parameters and serum hormone level, and fertility status. RESULTS: All patients had received multiple testosterone undecanoate (NebidoR) injections at local clinic due to androgen deficiency symptoms combined with lower serum testosterone level. The median duration of androgen replacement therapy prior to the development of azoospermia was 8 months (range: 4-12 months). After withdrawal of androgen therapy, sperm concentration and serum follicle-stimulating hormone level returned to normal range at a median 8.5 months (range: 7-10 months). CONCLUSION: Misusage of external androgen replacement therapy in infertile men with poor sexual function can cause temporary spermatogenic dysfunction, thus aggravating infertility.

Efficacy of In Vitro Germ Cell Culture in Nonobstructive Azoospermic Patients with Sertoli Cell Only Syndrome

PURPOSE: We determined the usefulness of in vitro germ cell culture in nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome, no sperm in testicular sperm extraction. MATERIALS AND METHODS: This study included 44 patients (45 testicular tissues) with nonobstructive azoospermia who were diagnosed with Sertoli cell only syndrome and were found to have no sperm in testicular sperm extraction between January 2006 and July 2008. Among the 45 testicular tissues, 22 tissues were processed for culture. In the in vitro cultures, the testicular tissues were dissociated and plated on gelatin-coated dishes. Patients were divided into 2 groups according to culture success: group I, culture positive (+; n=10); and group II, culture negative (-; n=12). RESULTS: The mean patient ages were 31.73 and 31.68 years for groups I and II, respectively. The mean testicular sizes were 10.19 and 10.42 cc, respectively; the semen volumes were 2.86 and 3.04 cc, respectively; and the mean FSH, LH, and testosterone levels were 18.86 mIU/ml, 5.99 mIU/ml, and 4.46 ng/ml vs. 21.02 mIU/ml, 6.29 mIU/ml, and 4.32 ng/ml for groups I and II, respectively, with no significant differences between the groups (p>0.05). The culture rate of nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome was 45.5% (10/22). Round spermatid injection was done in 2 patients with consent of the patients, but implantation failed. Among the 45 tissues, germ cells were found in 8 tissues after pathologic reexamination. CONCLUSIONS: The in vitro culture of germ cells would be useful in the advanced treatment of nonobstructive azoospermic patients.