[ effects of PLLA nanofiber scaffold on proliferation of frozen-thawed neonate mouse spermatogonial stem cells]

To investigate of the effects of a poly L-lactic acid [PLLA] nanofiber scaffold on proliferation of frozen-thawed neonate mouse spermatogonial stem cells. Spermatogonial cells were isolated from neonatal 3-6-day-old NMRI mice testes by two steps enzymatic digestion and differential plating. The isolated spermatogonial cells were divided into four culture groups: 1] fresh spermatogonial cells, 2] fresh spermatogonial cells seeded onto PLLA 3] frozen-thawed spermatogonial cells, 4] frozen-thawed spermatogonial cells seeded onto PLLA. Cells in all groups were cultured in DMEM supplemented with 5% FCS and 10 ng/ml GDNF for 3 weeks. Diameter and number of clusters which were determined during the culture and semi-quantitative RT-PCR were carried out at the end of 3rd week for all culture groups. Presence of spermatogonia at the culture was determined by reverse transcription polymerase chain reaction [RT-PCR] for several important spermatogonial markers [PLZF, Oct4, GFRalpha-1, VASA, ITGA6 and ITGB1]. The significancy of the data was analyzed using Repeated Measures and ANOVA tests. The findings indicated that the viability rate of the fresh cell [control 1 and exprimental 1] and the frozen cells after thawing [control 2 and exprimental 2] were 89.25 +/- 2.2 and 63 +/- 3.56, respectively and the differences were significant [p<0.001]. In vitro culturing of spermatogonial cells on PLLA significantly increased the formation of cell clusters in comparison with those of the control groups [p

[ effects of using retinoic acid during pregnancy on spermatogenesis and steroidogenesis in mature male rat]

The aim of this study was to investigate the effects of using Retinoic Acid during pregnancy on the spermatogenesis, testosterone and gonadotropin hormones in mature male rat. Pregnant rats were injected by 10, 20 and 30 mg/kg Retinoic acid [all-trans Retinoic Acid] intraperitoneally on the 8.5, 10.5, 12.5 and 14.5 days after copulation. An equivalent amount of the vehicle was similarly injected to corresponding control rats. Then animals did child birth. On time of maturity [10 weeks], from male animals in all of groups blood to determined the amount of hormones and the reproductive organs were separated. Morphometerical and Histological changes were studied in epididym and testis among experimental and control groups. The results were evaluated by using one way ANOVA and Tukey- test. Results indicateed that there were significant decrease in LH and FSH hormones, seminiferous tubules diameter, number of spermatocyte, spermatid and leydig cells experimental groups compared with the control group especially in animals which received 30 mg/kg of retinoic acid [p<0.05]. Also RA is the cause of decrease of the epididymis and deferent weight, and sperms in the epididymis, significantly [p<0.05]. Using of Retinoic Acid during pregnancy decreases the normal spermatogenesis and Steroidogenesis in mature male rats

[ case report: a rare anatomical variant of ansa cervicalis]

A rare variant of ansa cervicalis was discovered during routine dissection of the neck in a male cadaver. The superior root of the Ansa cervicalis was formed by the C1 ventral ramus. It fused with the vagus nerve in place of the hypoglossal nerve without any connection and descended into the carotid sheath with it. Then, C1 ventral ramus formed from the sheath and joined the inferior root from C2 and C3 ventral rami. We have reviewed the previous variation reports and reported a rare ansa cervicalis variation

[ effect of laminin and gelatin extracellular matrix on short-term cultivation of neonate mouse spermatogonial stem cells]

To compare the effect of laminin and gelatin on short-term culture of spermatogonial stem cells [SSCs] from neonatal mouse testes. Cell suspension containing SSCs were isolated from testes of 6 day-old mice and cultured in the presence of Glial-derived neuroterophic factor [GDNF], Epidermal Growth Factor [EGF] and Basic Fibroblastic Growth Factor [bFGF] on laminin-and gelatin-coated plates for 9 days. Number and area of colonies were measured in 5th, 7th and 9th days after culturing. At 9th day Immunostaining was used to detect expression of SSC markers, alpha 6-Integrin and beta 1-Integrin. moreover, the colonies were harvested and the percentage of alpha 6-Integrin and beta 1-Integrin positive cells was assessed by flowcytometery in both groups. Immunostaining analysis showed that our culture system contained SSC colonies as they were positive for alpha 6-Integrin and beta 1-Integrin. Additionally, the number of colonies those were formed on laminin were significantly higher in comparison with those of other group. but colony area was higher on gelatin. There was no significant difference in percentage of cells that expressed alpha 6-Integrin, beta 1-Integrin detected by flowcytometry in both groups. laminin as extracellular matrix cause to increase the number of neonate spermatogonial colonies and decrease the area of them [P