Mesenchymal Stem Cell Therapy of Male Infertility

Nowadays mesenchymal stem cell (MSC) therapy offers a broad spectrum of treatment of different conditions, including male infertility. Lots of studies suggest that injection of MSC promotes differentiation of germ cells and/or stimulates gonadal tissue development. Currently, there are plenty of MSC therapies of azoospermia which have been studied on animal models and demonstrated good results. Most of the studies were conducted using MSC derived from adipose tissue, the bone marrow, and umbilical cord. Despite the fact that this type of treatment in humans is not established as a first choice option, the use of these techniques gives us hope for a gradual introduction into our daily practice

One-Humped Camels (Camelus dromedaries) Hard Ticks Infestation in Qeshm Island, Iran

The economic importance of tick infestation on camels are important as they are important meat and milk producer animals in the less vegetation area of Iran and their health and production are greatly affected by the high tick infestation. In this investigation, tick infestations on camels (Camelus dromedarius) were determined in Qeshm Island, Iran. A total number of 912 adult ticks (472 males and 440 females) were collected and identified. Hyalomma dromedarii was the predominant tick specie and accounted for 61.9% of the adult ticks. Other hard ticks were H. anatolicum excavatum (22 %), H. asiaticum asiaticum (14.2 %), H. marginatum (1.9 %), H. impeltatum (0.4 %) and Ripicephalus bursa (0.4 %). In conclusion, The provision of tick control programs in the Qeshm Island would seem a prerequisite for improving camel meat and milk production

Effects of endurance exercise and estrogen supplementation on the proliferation of satellite cells

Abstract Animal and human studies indicated that overtension and stress release inflammatory substances and growth factors that are produced following exercise, which leads to satellite cell activation and proliferation. The aim of the present study was to investigate the effect of an 8-week endurance exercise and estrogen supplementation on the proliferation of satellite cells in rats. Seventy-six rats were selected and randomly divided into two equal groups, ovariectomized and intact groups. Both groups were randomly divided into four subgroups as follows: endurance exercise, estrogen supplementation, estrogen supplementation with endurance exercise, and control. Then, the endurance exercise group and estrogen supplementation with endurance exercise group performed endurance exercise for 8 weeks, three sessions per week. In each week, the estrogen supplementation group and estrogen supplementation with endurance exercise group were injected subcutaneously with 3 mg/kg of estradiol benzoate. The soleus muscle was retracted and placed into 10 % buffered formalin solution. In a pathological lab, the number of satellite cells was counted and recorded using a light microscope through hematoxylin and eosin staining and immunohistochemistry for CD56. Increase in satellite cell number was significant in the two groups of intact rats treated with estrogen supplementation and the ovariectomized rats which performed endurance exercise. The comparison of these groups' means demonstrated that the satellite cell number increased more in the ovariectomized rats. Endurance exercise and estrogen supplementation can increase the proliferation of satellite cells in the rat's soleus muscle

Growth kinetics and characterization of human dental pulp stem cells: comparison between third molar and first premolar teeth

Dental pulp stem cells (DPSCs) play an important role in tissue regeneration. This study compares the growth kinetics and characterization of third molar and first premolar human DPSCs. Dental pulp tissues were isolated from human first premolar and third molar teeth and were digested by treating them with collagenase type I. Single-cell suspensions from each dental pulp were seeded in T25 culture flasks and the media were replaced every 3 days until 70% confluence. The cells were enumerated to determine the population doubling time (PDT). Cells were characterized using flow cytometry, RT-PCR and osteogenic medium for differentiation of DPSCs. Karyotyping assay was also performed till passage 7th. The DPSCs had spindle-shaped morphology. There was an increase in PDT in third molar DPSCs when compared to first premolar teeth. Positive expression of CD44, CD73, and CD90 and negative expression of CD34 and CD45 were illustrated. A normal karyotype was visible for all seven passages. The Alizarin red staining was positive for osteogenic induction of DPSCs. When DPSCs are needed, third molar teeth can be a good and convenient candidate for cell transplantation, yielding high number of cells with mesenchymal characteristics. They can be a source for further investigations in vitro and work on tissue engineering protocols

Maternal aromatase inhibition via letrozole altered RFamide-related peptide-3 and gonadotropin-releasing hormone expression in pubertal female rats

Objective(s): Despite prevalence of polycystic ovary syndrome (PCOS) among childbearing women and development of many animal models for this syndrome, information on its etiology is still scarce. The intrauterine hyperandrogenic environment may underlie changes at the level of hypothalamus, pituitary, ovary organization in female offspring, and PCOS later in life. Letrozole has been shown to mimic reproductive and metabolic characteristics of PCOS in adult rodent models. Therefore, this research aimed to assess the condition in a prenatal letrozole-treated rat model. Materials and Methods: Twenty-eight female rats dams receiving letrozole at certain doses during late pregnancy were used in the trial. Pregnant Sprague-Dawley rats (n=21) received letrozole treatment on gestation days 16–18 at doses of 1.25, 1.0, 0.75, 0.5, and 0.25 mg/kg body weight (BW). Results: Prenatal letrozole treatment delayed parturition time and reduced the litter size in pregnant dams (P<0.0001). Late puberty onset, irregular ovarian cyclicity, increased anogenital distance (AGD), body weight gain, serum testosterone concentration, and reduced estradiol levels (P<0.0001) were observed in the female offspring of dams receiving 1.25 and 1 mg/kg BW letrozole. Furthermore, letrozole at 1.25 and 1 mg/kg BW showed increased RFRP and decreased GnRH mRNA expression (P<0.0001). Letrozole treatment at doses of 1 mg/kg BW and lower was not fetotoxic. Conclusion: It was concluded that 1 mg/kg BW letrozole may be suggested for prenatal PCOS induction. Keywords Gonadotropin-releasing hormone Hypothalamus Letrozole Polycystic ovary syndrome Prenatal Rat RFamide-related peptide-

Promjene u RF-amidu srodnom peptidu-3 hipotalamusa i ekspresijama gena Kiss1 tijekom spermatogeneze kod štakora u uvjetima kroničnog stresa.

The effects were evaluated of chronic stress and the glucocorticoid receptor antagonist (RU486) on mRNA expressions of RF-amide related peptide-3 (RFRP-3) in the dorsomedial hypothalamic nucleus (DMH) and Kiss1 in the arcuate nucleus (ARC) of male rats. Twenty-four male rats were allocated to four equal sized groups: the stress, RU486, stress/RU486, and control groups. In the stress group the rats were restrained 1 hour/day for 12 days. In the RU486 group, the rats were injected with RU486 for 12 days. In the stress/RU486 group, the rats were injected with RU486 1 hour before the stress process for 12 days. Relative expressions of RFRP-3 and Kiss1 mRNAs were determined using real-time PCR. The relative expression of RFRP-3 mRNA in the stress group was higher than that in the RU486 and control rats. The relative expression of RFRP-3 mRNA did not differ between the stress group and the stress/RU486 rats. Furthermore, the relative expressions of Kiss1 mRNA in the stress, RU486, and stress/RU486 groups were less than that of the control rats. The relative expression of Kiss1 mRNA did not differ between the stress, RU486, and stress/RU486 groups. In conclusion, dysfunction in male rat fertility caused by the chronic stress may be the result of the increase in REFP-3 and the decrease in Kiss1 mRNA expression.Istražen je učinak kroničnog stresa i antagonista glukokortikoidnog receptora (RU486) na ekspresiju mRNA RF-amidu srodnog peptida-3 (RFRP-3) u dorzomedijalnoj jezgri hipotalamusa (DMH), te na ekspresije gena Kiss1 u arkuatnom nukleusu (ARC) štakora. Dvadeset i četiri štakora bila su raspodijeljena u četiri jednake skupine: stresna skupina, RU486 skupina, stresna/RU486 skupina i kontrolna skupina. U stresnoj skupini štakori su 12 dana bili obuzdani tijekom jednog sata dnevno. U skupini RU486, štakorima je tijekom 12 dana bio primijenjivan RU486. U skupini stres/RU486, štakorima je tijekom 12 dana apliciran RU486 jedan sat prije postupka obuzdavanja. Relativne ekspresije RFRP-3 i Kiss1 mRNA određene su lančanom reakcijom polimerazom u stvarnom vremenu. Relativna ekspresija RFRP-3 mRNA u stresnoj skupini bila je veća nego u skupini RU486 i kontrolnoj skupini. Relativna ekspresija RFRP-3 mRNA nije bila različita između stresne skupine i stres/RU486 skupine. Nadalje, relativne ekspresije Kiss1 mRNA u stresnoj skupini, skupini RU486, i stresnoj skupini/RU486 bile su manje u odnosu na kontrolnu skupinu. Relativna ekspresija Kiss1 mRNA nije se razlikovala između stresne skupine, skupineRU486 i stresne skupine/RU486. Zaključno, disfunkcija plodnosti kod štakora izloženih kroničnom stresu može biti uzrokovana putem povećane ekspresije RFRP-3 i smanjene ekspresije Kiss1 mRNA

Caprine Endometrial Mesenchymal Stromal Stem Cell: Multilineage Potential, Characterization, and Growth Kinetics in Breeding and Anestrous Stages

The endometrial layer of the uterus contains a population of cells with similar characteristics of mesenchymal stem cells (MSCs). In the present study, caprine endometrial mesenchymal stromal stem cells (En-MSCs) characters and differentiation potential to chondrogenic, osteogenic, and adipogenic cell lines as well as their growth kinetics in breeding and anestrous stages were evaluated. En-MSCs were enzymatically isolated from endometrial layer of the uterus of adult goats and were cultured and subcultured until passage 4. The growth kinetics and population doubling time (PDT) of caprine En-MSCs in breeding and anestrous stages were determined. En-MSCs in passage 4 were used for the karyotyping and differentiation into chondrocytes, osteocytes, and adipocytes. The PDT in anestrus phase was 40.6 h and in cyclic goats was 53 h. En-MSCs were fibroblast-like in all passages. The number of chromosomes was normal (2n=60) with no chromosomal instability. Chondrogenic, osteogenic, and adipogenic differentiation of En-MSCs was confirmed by staining with Alcian blue, Alizarin red, and Oil Red O, respectively. Caprine En-MSCs demonstrated to be an alternative source of MSCs for cell therapy purposes in regenerative medicine

Ekspresija mRNA agutiju srodnog peptida i mRNA melanokortin-4 receptora u arkuatnoj jezgri za vrijeme gravidnosti i laktacije štakorica.

Pregnancy is associated with a range of physiological adjustments to adapt the body to the demands of the growth of the fetus and subsequent lactation. It has been observed that agouti-related peptide (AGRP) and melanocortin-4 receptor (MC4R) are involved in energy homeostasis. A randomized controlled experimental study was planned to investigate the expression of AGRP and MC4R mRNAs in the stages of pregnancy and lactation in rat arcuate nucleus (ARC) of the hypothalamus. Thirty-two adult female rats were randomly divided into six groups. Pregnant rats were assigned into three groups (n = 6) of 7, 14, and 21 days of pregnancy. Two more groups were also assigned of non-suckling rats (n = 5), immediately separated from their pups after parturition, and suckling rats (n = 5), allowed to suckle five pups until day 8 (increasing milk). The sixth group consisted of four ovariectomized rats, which were assigned two weeks after surgery and served as control. Using real-time PCR, the relative expressions (compared to controls) of MC4R and AGRP mRNAs in ARC were calculated in the pregnant, suckling and non-suckling rats. Expression of AGRP mRNAs in pregnant rats on days 14 and 21 was higher than that observed in suckling and non-suckling rats (P<0.05). Expression of MC4R mRNAs in pregnant rats on days 14 and 21 was lower than that observed in suckling rats, and was higher on day 7 than that observed in both suckling and non-suckling rats (P<0.05). In conclusion, expression of AGRP in pregnancy and MC4R in lactation in ARC of rats controls energy homeostasis.Gravidnost je povezana s nizom fizioloških prilagodbi kojima tijelo odgovara zahtjevima povezanima s rastom fetusa i kasnije laktacije. Uočeno je da su agutiju srodan peptid (AGRP) i melanokortin-4 receptor (MC4R) uključeni u energetsku homeostazu. Randomiziranim kontroliranim pokusom planirano je u arkuatnoj jezgri (ARC) hipotalamusa štakorica istražiti ekspresiju mRNA AGRP i mRNA MC4R tijekom gravidnosti i laktacije. Trideset dvije odrasle štakorice nasumično su bile podijeljene u šest skupina. Gravidne su štakorice podijeljene u tri skupine (n = 6), s obzirom na 7., 14. i 21. dan gravidnosti. Još dvije skupine (n = 5) činile su nedojne štakorice koje su odvojene od svoje mladunčadi odmah nakon porođaja te dojne štakorice kojima je dozvoljeno dojenje 5 mladunaca do osmog dana rastuće laktacije. Četiri ovarijektomizirane štakorice, dva tjedna nakon operacije, dodijeljene su u 6. kontrolnu skupinu. Koristeći PCR u stvarnom vremenu, relativna ekspresija (usporedba s kontrolama) mRNA MC4R i mRNA AGRP u ARC izračunata je za gravidne, nedojne i dojne štakorice. Ekspresija mRNA AGRP kod gravidnih štakorica 14. i 21. dan bila je veća od one opažene kod dojnih i nedojnih štakorica (P<0,05). Ekspresija MC4R mRNA u gravidnih štakorica 14. i 21. dan bila je manja u odnosu na dojne štakorica i veća 7. dan u odnosu na skupine dojnih i nedojnih štakorica (P<0,05). Zaključno, ekspresija AGRP u ARC tijekom gravidnosti i MC4R u ARC tijekom laktacije kontrolira energetsku homeostazu štakorica

Establishment, Culture, and Characterization of Guinea Pig Fetal Fibroblast Cell

Establishment of Guinea pig fetal fibroblast cells and their biological evaluation before and after cryopreservation were the main purposes of this study. After determination of the proper age of pregnancy by ultrasonography, 30 days old fetuses of Guinea pigs were recovered. Their skins were cut into small pieces (1 mm2) and were cultured. When reaching 80–90% confluence, the cells were passaged. Cells of the second and eighth passages were cultured in 24-well plates (4×104 cells/well) for 6 days and three wells per day were counted. The average cell counts at each time point were then plotted against time and the population doubling time (PDT) was determined. Then, vials of cells (2×106 cells/mL) were cryopreserved for 1 month and after thawing, the cell viability was evaluated. The PDT of the second passage was about 23 h and for the eighth passage was about 30 h. The viability of the cultures was 95% in the second passage and 74.5% in the eighth passage. It was shown that the Guinea pig fetal fibroblast cell culture can be established using the adherent culture method while, after freezing, the viability indices of these cells were favorable