epidermal-growth-factor and Infertility--Male

Orchiopexy is one of the most frequently used surgical procedures for cryptorchidism and has been shown to have a beneficial effect on fertility. However, orchiopexy, especially for bilateral cryptorchidism, does not always guarantee subsequent fertility and paternity. Compared with a control group, paternity was significantly compromised in men with previous bilateral, but not unilateral cryptorchidism. Recent techniques of assisted reproductive technology, especially testicular sperm extraction with intracytoplasmic sperm injection (TESE-ICSI), have brought revolutionary changes in clinical therapy for infertiliy. If spermatozoa exists in testis of infertile men, logically there is a possibility of paternity. However, our study demonstrated that about 20% of pubertal boys who had had orchiopexy, were predicted to have lost their future paternity potential even if TESE-ICSI were conducted, because they were predicted to have no spermatozoa in the testis. To prevent or reverse the damage of spermatogenesis at prepuberty or puberty, we should not take a wait-and-see attitude but should consider a countermeasure for the pubertal boys who had had bilateral orchiopexy in childhood, especially when the serum follicle stimulating hormone level is elevated and testicular volume is lowered, before paternity is lost. In this review, we discuss the potential approaches including epidermal growth facter therapy, gene therapy and stem-cell therapy for cryptorchid patients in the future.

Topics: Adolescent; Age Factors; Animals; Child; Cryptorchidism; DAX-1 Orphan Nuclear Receptor; DNA-Binding Proteins; Embryonic Stem Cells; Epidermal Growth Factor; Fertility; Genetic Therapy; Humans; Infertility, Male; Male; Puberty; Receptors, Retinoic Acid; Repressor Proteins; Reproductive Techniques, Assisted; Spermatogenesis; Urologic Surgical Procedures, Male

Topics: Epidermal Growth Factor; Genetic Predisposition to Disease; Genotype; Humans; Infertility, Male; Male; Polymorphism, Genetic; Polymorphism, Single Nucleotide

To study the effects of serum and growth factors on propagation of porcine male germline stem cells (MGSCs) in vitro and develop a culture system for these stem cells.. Fresh testicular cells from neonatal piglets were obtained by mechanical dissociation and collagenase-trypsin digestion. After differential plating, non-adhering cells were cultured in media supplemented with different concentrations of serum (0, 1 %, 2 %, 5 %, 10 %). After 10 days of primary culture, the cells were maintained in media supplemented with different concentrations of growth factors (basic fibroblast growth factor and epidermal growth factor at 1, 5, 10 ng/ml). The number of MGSC-derived colonies with different sizes was determined in each treatment to assess the effects of serum concentrations and growth factors.. The number of MGSC-derived colonies was significantly higher in the presence of 1 % rather than 10 % fetal bovine serum (FBS). Basic fibroblast growth factor (bFGF) at 1, 5 ng/ml and epidermal growth factor (EGF) at 5, 10 ng/ml significantly promoted colony formation. Immunocytochemistry, reverse transcriptase-polymerase chain reaction (RT-PCR) and xenotransplantation assays demonstrated the presence of functional stem cells in cultured cell population.. In vitro propagation of porcine MGSCs could be maintained in the presence of 1 % FBS and supplementation of growth factors for 1 month.

Topics: Adult Stem Cells; Alkaline Phosphatase; Animals; Cell Culture Techniques; Cell Proliferation; Culture Media; Epidermal Growth Factor; Fibroblast Growth Factor 2; Infertility, Male; Male; Mice; Mice, Inbred BALB C; Spermatogenesis; Spermatozoa; Swine; Testis; Transplantation, Heterologous

Human cloned blastocysts generated from oocytes following in vitro maturation (IVM) are a potential resource for embryonic stem cells (ESC) with homologous immune systems. The purpose of this study was to evaluate the effects of multiple growth factors [epidermal growth factor (EGF), brain-derived neurotrophic factor (BDNF) and insulin-like growth factor-1 (IGF-1)] on human oocyte maturation, early embryo development, blastocyst formation and ESC line generation.. Patients (n= 344) undergoing IVF owing to male factor infertility were enrolled in this study. Metaphase II oocytes were separated into four grades based on their morphology. Spindle assembly from IVM oocytes with or without growth factor treatment was assessed by immunostaining. Piezo-assisted micromanipulation technology was used to produce fertilized (ICSI) and cloned [(somatic cell nuclear transfer (SCNT)] embryos. Embryos received four different growth factor treatments; embryo development rates from pronuclear to blastocyst stage and embryo grading (for quality) at the 8-cell stage were analyzed. The presence of receptors on human cumulus cells and IVM oocytes was assessed by immunofluorescence. The blastocysts generated from fertilized and cloned embryos were used for ESC derivation.. The combination of EGF, BDNF and IGF-1 can effectively increase oocyte maturation rate in vitro, and significantly improve the oocyte quality in terms of morphology and normal spindle levels (P< 0.05). Also, the developmental competence of fertilized oocytes to 8-cell and blastocyst stages was improved by the addition of growth factors (P< 0.05). However, there were no significant differences among the four groups in 8-cell grading. Blastocyst formation in cloned embryos cultured with the three growth factors was higher than the control group (23.1 versus 4.3%, P< 0.05). Receptors for the three growth factors were present in cumulus cells and IVM oocytes, and four human ESC lines were derived from fertilized blastocysts but none from cloned blastocysts.. This study demonstrated that EGF, BDNF and IGF-1 can improve oocyte maturation rate and quality in vitro, and consequently increase early embryo development and blastocyst formation, which is very beneficial in improving the reprogramming efficiency of SCNT. The present study has identified a valuable culture system for IVM and cloned human embryos, potentially using these embryos to derive human therapeutic ESC.

Topics: Blastocyst; Brain-Derived Neurotrophic Factor; Cloning, Organism; Cumulus Cells; Embryonic Development; Embryonic Stem Cells; Epidermal Growth Factor; Female; Fertilization; Fertilization in Vitro; Fibroblasts; Humans; Infertility, Male; Insulin-Like Growth Factor I; Karyotyping; Male; Metaphase; Microscopy, Confocal; Microscopy, Fluorescence; Oocytes; Sperm Injections, Intracytoplasmic

This study aims to proliferate spermatogonial stem cells (SSCs) and compare the in-vitro effects of laminin and growth factors on the proliferation of adult human SSC.. Isolated testicular cells were cultured in DMEM supplemented with 5 % fetal calf serum (FCS). During the culture, enriched spermatogonial cells were treated with a combination of glial cell line-derived neurotrophic factor (GDNF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and mouse leukemia inhibitory factor (LIF) in the presence or absence of human placental laminin-coated dishes. Cluster assay was performed during culture. Presence of spermatogonia was determined by an ultrastructural study of the cell clusters, reverse transcription polymerase chain reaction (RT-PCR) for spermatogonial markers and xenotransplantation to the testes of busulfan-treated recipient mice. Statistical significance between mean values was determined using statistical ANOVA tests.. The findings indicated that the addition of GDNF, bFGF, EGF and LIF on laminin-coated dishes significantly increased in-vitro spermatogonial cell cluster formation in comparison with the control group (p ≤ 0.001). The expression of spermatogonial markers was maintained throughout the culture period. Furthermore, a transplantation experiment showed the presence of SSC among the cultured cells. In addition, a transmission electron microscopy (TEM) study suggested the presence of spermatogonial cells of typical morphology among the cluster cells.. It can be concluded that human SSCs obtained from non-obstructive azoospermic (NOA) patients had the ability to self-renew in the culture system. This system can be used for the propagation of a small number of these cells from small biopsies.

Topics: Analysis of Variance; Animals; Azoospermia; Biomarkers; Biopsy; Busulfan; Cell Count; Cell Culture Techniques; Cell Proliferation; Cells, Cultured; Culture Media; Epidermal Growth Factor; Fibroblast Growth Factor 2; Glial Cell Line-Derived Neurotrophic Factor; Humans; Infertility, Male; Laminin; Leukemia Inhibitory Factor; Male; Mice; Microscopy, Electron, Transmission; Reverse Transcriptase Polymerase Chain Reaction; Serum; Spermatogonia; Stem Cell Transplantation; Stem Cells; Testis; Transplantation, Heterologous

Detection of EGF and IGF-I in human embryo cultures and their effect on ICSI outcome.. Collection of culture medium from embryos of 50 women under ICSI program. EGF and IGF-I were measured via enzyme immunoassay.. ICSI outcome was independent of age, infertility years, FSH, LH, prolactine and E2. EGF detection was higher in 48- (32%), than in 72-hour embryos (14%) (p < 0.001). EGF negative embryos are likely to be arrested at the morula stage (p < 0.001) and are associated with poor pregnancy rates (p < 0.05). IGF-I was undetected in 48-hour embryos.. For the first time human embryos were surveyed from fertilization until embryo transfer, regarding EGF and IGF-I production. IGF-I is not a predictor of ICSI outcome. EGF is present in one-third of human embryo cultures at 48 hours, but this ratio wanes at the morula stage. EGF negative embryos are associated with lower pregnancy rates.

Topics: Adult; Age Factors; Blastocyst; Cleavage Stage, Ovum; Culture Media; Embryo Implantation; Embryo Transfer; Epidermal Growth Factor; Extraembryonic Membranes; Female; Humans; Infertility, Male; Insulin-Like Growth Factor I; Male; Morula; Pregnancy; Pregnancy Rate; Risk Factors; Sperm Injections, Intracytoplasmic; Time Factors

The relationship between male reproductive function and the blood plasma level of epidermal growth factor (EGF) is of interest in the light of the role that circulating EGF appears to play in regulating mouse spermatogenesis. We measured the concentrations of EGF in the blood plasma of 39 fertile men (sperm count > 20 x 10(6)/ml) and compared them with those of 31 infertile men (sperm < 20 x 10(6)/ml). Blood plasma levels of follicle stimulating hormone (FSH), luiteinising hormone (LH), prolactin and testosterone were also determined. The infertile patients had mean blood plasma EGF concentrations of 0.75 +/- 0.10 ug/L. The value was significantly lower than that of the fertile group (1.28 +/- 0.14 ug/L; P < 0.005). There were statistically significant differences between the fertile and infertile groups in sperm count, sperm viability, mean forward progression, testosterone, LH and FSH (P values between 0.0001 and 0.023). There was no significant difference in the prolactin concentrations between the two groups. Although overall average blood plasma EGF concentrations are significantly lower in the infertile males, regression analysis failed to reveal any direct relationships among the various parameters studied.

Topics: Adult; Analysis of Variance; Case-Control Studies; Epidermal Growth Factor; Follicle Stimulating Hormone; Humans; Infertility, Male; Luteinizing Hormone; Male; Middle Aged; Prolactin; Regression Analysis; Sperm Count; Sperm Motility; Spermatogenesis; Testosterone

We analyzed expression of transforming growth factor (TGF)-alpha, epidermal growth factor (EGF) and their receptor, EGF receptor (EGFR), by immunohistochemistry in the human testis to determine the possible roles of these growth factors in human testicular function. Specimens were obtained from 17 patients including 9 patients with infertility, 4 patients with prostatic carcinoma and 4 patients with contralateral testicular tumor. EGF immunoreactivity was positive in the hyperplasic Leydig cells of one patient but negative in the other cases. On the other hand, strong TGF-alpha immunoreactivity was observed in Leydig cells, with weak staining in Sertoli cells and germ cells in cases with normal spermatogenesis. EGFR immunoreactivity was observed in the Leydig and peritubular cells, appearing as membrane staining. Marked immunoreactivity for TGF-alpha was observed in the Sertoli cells in testes with decreased spermatogenesis, especially in the Sertoli-cell-only syndrome. This finding may indicate a compensatory increase of TGF-alpha expression in the Sertoli cells accompanying a decrease in spermatogenesis. No significant correlation was found between the degrees of spermatogenesis and immunolocalization of the EGF receptor. These findings suggest that TGF-alpha is a locally produced growth factor that is involved in spermatogenesis in the human testis via an autocrine and/or paracrine mechanism.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Epidermal Growth Factor; ErbB Receptors; Humans; Immunohistochemistry; Infertility, Male; Male; Middle Aged; Orchiectomy; Prostatic Neoplasms; Spermatogenesis; Testicular Neoplasms; Testis; Transforming Growth Factor alpha

We measured immunoreactive epidermal growth factor (EGF) by a homologous RIA in seminal plasma (SP) from 31 fertile and 52 infertile men to determine the relationship between SP EGF levels and total sperm count in the ejaculates. The mean SP EGF levels in fertile and infertile men were 41.7 +/- 21.5 (+/- SD) and 53.1 +/- 30.8 micrograms/L, respectively. Infertile men with sperm-associated immunoglobulin G (n = 9), immunoglobulin A (n = 6), or both (n = 8) had mean SP EGF levels of 48.9 +/- 26.1, 47.9 +/- 17.5, and 56.5 +/- 32.1 micrograms/L, respectively. Seven men with severe oligospermia had a mean SP EGF level of 58.5 +/- 35.9 micrograms/L. There was no correlation (r = 0.14; P greater than 0.05) between SP EGF levels and total sperm counts in these men. Fractionation of SP by high performance liquid chromatography on a size exclusion (TSK G2000 SW) column revealed a single immunoreactive peak with an approximate mol wt of 8000, slightly higher than the mol wt of circulating human EGF (6000). We conclude that SP EGF may be distinct from peripheral plasma EGF.

Topics: Adult; Antibodies; Chromatography, Gel; Chromatography, High Pressure Liquid; Epidermal Growth Factor; Histocytochemistry; Humans; Immunoglobulin G; Infertility, Male; Male; Oligospermia; Radioimmunoassay; Semen; Sperm Count; Staining and Labeling

Epidermal growth factor/urogastrone (EGF/URO) was measured in the seminal plasma of 214 untreated patients attending an infertility clinic. Patients were assigned groups according to sperm density and progressive motility and in the case of azoospermia into an obstructive etiology or germinal failure. There was no significant variation in mean EGF/URO levels between groups, suggesting that this peptide plays no role in the density or motility of sperm associated with fertility. Other functions this peptide may fulfill in the female reproductive tract, including that of immunosuppression, are discussed.

Topics: Epidermal Growth Factor; Humans; Infertility, Male; Male; Semen; Sperm Count; Sperm Motility

In the present study, we have partially purified a characterized epidermal growth factor (EGF)-like substance(s) from human seminal plasma, and determined the concentrations of immunoreactive (IR)-hEGF in seminal plasma from normal and infertile males. Competitive binding curves of seminal plasma extracts were parallel to those of standard hEGF in both radioimmunoassay and receptor assay. Seminal IR-hEGF was similar to standard hEGF by gel exclusion chromatography, isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The concentrations of IR-hEGF in normal seminal plasma (48 +/- 9 ng/ml) did not differ from those of infertile males (41 +/- 3 ng/ml); the concentrations of seminal plasma IR-hEGF did not correlate with density, motility or morphology of sperm. These data clearly demonstrate the presence of hEGF in human seminal plasma indistinguishable from hEGF of urinary origin, and suggest that it may not play an important role in the sperm function. The tissue(s) of its origin and its physiological function in the male reproductive organs remain undetermined.

Topics: Adult; Epidermal Growth Factor; Humans; Infertility, Male; Male; Radioimmunoassay; Radioligand Assay; Reference Values; Semen