Xeno-free culture of human spermatogonial stem cells supported by human embryonic stem cell-derived fibroblast-like cells.

Spermatogonial stem cells (SSCs) divide continuously to support spermatogenesis throughout postnatal life and transmit genetic information to the next generation. Here, we report the successful establishment of the method for the isolation and identification of human SSCs from testicular tissue, and to determine the culture conditions required to expand SSCs on human embryonic stem cell-derived fibroblast-like cells (hdFs). Large-scale cultures of SSCs were maintained on hdF feeder layers and expanded in the presence of a combination of cytokines and glial cell line-derived neurotrophic factor for at least 2 months.

[The feeder layer of human embryonic fibroblasts supports the growth of human spermatogonial stem cells].

Objective: To investigate the methods and conditions for the isolation, purification and culture of human spermatogonial stem cells (SSCs) on the feeder layer cells of human embryonic fibroblasts (hEFs).

Methods: SSCs isolated and purified from normal human fetal testicular tissues by sequential two-step enzyme digestion and Percoll uncontinuous density gradient centrifugation were cultured on the feeder layer cells of hEFs isolated from 5-9 weeks old human embryos. The surface markers SSEA-1 and OCT4 of the SSCs were detected by immunohistochemistry; the alkaline phosphatase (AKP) activity of the SSC clones measured; and the expressions of the SSC-related genes determined by RT-PCR.

Leptin and varicocele-related spermatogenesis dysfunction: animal experiment and clinical study.

The objective of this study was to explore the relationships between varicocele-related spermatogenesis dysfunction and the expression of leptin and leptin receptors. In rats with experimental varicocele, the function of spermatogenesis, the expression of leptin and leptin receptors in testes were analysed; and in patients with varicocele-related male infertility, serum and seminal plasma levels of leptin, gonadal hormones and semen parameters were evaluated. In the testes of rats, leptin was expressed in seminiferous tubules and intersitium, leptin receptor was predominantly expressed in interstitium.

[Effects of growth hormone supplementation on erectile function and expression of nNOS in aging rats].

Objective: To explore the effects of growth hormone( GH) supplementation on erectile function and expression of nNOS in the intracavernosal nerves in aging rats.

Methods: Twenty male Sprague-Dawley rats aged 18 months were randomly divided into Groups A and B, and ten 2-month-old male Sprague-Dawley rats included in Group C. 1 U/(kg x d) GH was given to Group A, and the same volume of saline to Groups B and C.

[Seminal plasma angiotensin II detection and its clinical implication].

Objective: To investigate the variation of seminal plasma angiotension II (Ang II) in infertile men and its clinical implication.

Methods: Ang II values in paired blood plasma and seminal plasma from 43 infertile men(13 azoospermia, 8 asthenozoopermia, 17 asthenozoospermia and 5 cases with normal semen parameters) and 10 normal controls were obtained by SPE-HPLC-RIA. All semen samples with spermatozoa were analyzed by CASA for sperm count, motility and other parameters.

[Experimental research on human spermatozoa membrane proteins by two-dimensional gel electrophoresis].

Objectives: To analyze human spermatozoa membrane proteins by two-dimensional gel electrophoresis and to provide a basis for drawing the protein map of normal human spermatozoa membrane proteins.

Methods: Spermatozoa were purified by Percoll density centrifugation, and spermatozoa membrane proteins were analyzed by two-dimensional gel electrophoresis using isoelectric focusing and polyacrylamide gel electrophoresis.

Results: About 800 protein spots could be identified by the imaging analysis system.

[Measurement of the reactive oxygen species and cytokines in the seminal plasma of leukocytospermic patients].

Objectives: To detect the levels of reactive oxygen species (ROS), superoxide dismutase(SOD) and interleukin 8(IL-8) in seminal plasma of infertile patients, and evaluate the possible relationship between those levels.

Methods: Semen was collected from normal donors (15 cases), infertile men without infection (16 cases), and infertile men with infection (leukocytospermia, 11 cases). The routine analysis of semen was accomplished, and then the levels of IL-8, malondialdehyde (MDA), SOD, and white blood cell (WBC) were examined.

Cryopreservation-induced decrease in heat-shock protein 90 in human spermatozoa and its mechanism.

Aim: To study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism.

Methods: In 18 healthy men, the seminal sperm motility and HSP90 levels were studied before and after cryopreservation using SDS-PAGE, Western blotting and computerized image analysis.

Results: The sperm motility declined significantly after cryopreservation (P<0.

[The effects of testosterone undecanoate on relaxation of rat corpus cavernosum smooth muscle in vitro].

Objectives: To investigate the role of Undecanoate (Andriol), as a kind of testosterone, in regulating the relaxation of isolated rat corpus cavernosum in vitro.

Methods: The castrated rats were given high and low dosage Andriol respectively, compared with intact and castrated rats. After treatment of 4 weeks, the corpora cavernosa were cut, trimmed as to strips.