مجله طب دامی ایران
سال:1392 | دوره:7 | شماره:2 |تعداد مقالات:14

BACKGROUND: Spermatogonial stem cells (SSCs) are infrequent self-renewing cells among the type A spermatogonia within the seminiferous tubules and are the basis of spermatogenesis in mammalian testis. An adequate number of SSCs is a primary requirement for the study of their behavior, regulation, and further biomanipulation.OBJECTIVES: In this paper, we studied the development of the primary co-cultures of type A spermatogonia and prepubertal bovine sertoli cells in the presence of Colony Stimulating Factor 1 (CSF1), a potential contributor in the SSC niche.METHODS: The effect of different concentrations of CSF1 (0, 10, 50 and 100 ng/mL) on the colonization activity of spermatogonial cells was assessed 4, 7 and 11 days after the beginning of the culture by counting the total number of colonies and measuring their area in each group of the present experiment. Immunofluorescent staining against OCT4 and vimentin led to the confirmation of the nature of both the SSCs and sertoli cells.RESULTS: Results showed that the total number of colonies from day 4 to 11 increased significantly in all groups, independent of CSF1 concentration. In addition, the total number and total area of colonies were higher (not significant) in 10 and 50 ng/mL CSF1 treatments than the control and 100 ng/mL CSF1 groups in all the three evaluations during the experiment. However, this difference was only significant (p<0.05) between the total area of colonies in the control and 10 ng/mLCSF1 groups at day 4 of co-culture.CONCLUSIONS: It was concluded that CSF1 can be a suitable growth factor for improving SSCs colonization in vitro, particularly during the first days of culture where accompanying sertoli cells still have not proliferated sufficiently to support the propagating spermatogonial cells.

BACKGROUND: Influenza outbreak has become a great lifethreatening disease in the world. Nasal vaccines can induce systemic IgG and mucosal IgA antibody responses, which establish two layers of immune defense against the infectious pathogens like influenza. Mucosal vaccines must overcome several limitations, including the mucociliary clearance and inefficient uptake of soluble antigens. Therefore, nasal vaccines require potent adjuvants and delivery systems.OBJECTIVES: In this study we evaluated the effect of N-trimethyl chitosan (TMC) as a potent vehicle for DNA encoding M2e/HSP70c in order for intranasal administration in mice.METHODS: Ectodomain of the conserved influenza matrix protein 2 (M2e), which has been found to induce heterosubtypic immunity, was fused to HSP70359-610 or C-terminus of Mycobacterium tuberculosis HSP70 (HSP70c) in pcDNA3.1 vector (pcDNA/M2e-HSP70c) and then encapsulated into a derivative of chitosan, N-trimethyl chitosan (TMC). After encapsulation of the plasmid, physical properties of the particles were investigated using ZetasizerÒ 3000 the particles were then administered through the intranasal delivery in BALB/c mice.RESULTS: It was found that the particles had a size ranging between 90-120nm and positive surface charge. The intranasal immunization with M2e- HSP70c+TMC in BALB/c mice significantly induced higher M2e specific IgG than those induced in control groups (pcDNA/M2e-HSP70c without TMC, pcDNA/M2e, bearing M2e alone, and PBS).CONCLUSIONS: The present study showed that the encapsulation of M2e/ HSP70c into N-trimethyl chitosan (TMC) could strongly induce the humoral immune response against the M2e-HSP70c plasmid without lowering the adjuvant efficacy of HSP70c.

BACKGROUND: Avian reoviruses (ARVs) are members of the Orthoreovirus genus; one of the 12 genera of the Reoviridae family. The ARVs are the cause of some important diseases in poultry such as reovirus-induced arthritis, tenosynovitis, chronic respiratory disease, and mal-absorption syndrome.OBJECTIVES: In this study, the presence of ARVs in the Iranian breeder flocks was investigated through reverse transcriptionpolymerase chain reaction (RT-PCR) and restriction enzyme fragment length polymorphism (RFLP).METHODS: A total of 800 fecal swab samples were initially collected from breeder flocks (older than 45 weeks of age). They were then sent to the laboratory in containers with PBS, and after that they were pooled and finally to 120 samples were obtained. The total RNA extracted from the pooled fecal samples were used to amplify the selected parts of the S1 (1023 bp) and S4 (437 bp) genes from the ARV field isolates using RT-PCR. The positive RT-PCR amplified products were further analyzed by RFLP using five restriction enzymes.RESULTS: Based on the findings, 5 samples were positive with the S1 primer and 6 samples were with the S4 one. The patterns observed after the digestion of PCR products revealed that the isolates of this study were identical to both the S1133 vaccine and standard strains.CONCLUSIONS: The findings suggested that the RT-PCR/RFLP analysis might be considered as a simple and rapid approach for the differentiation of ARVisolates. This study was the first molecular detection of the ARVs presence in the Iranian breeder flocks using the RTPCR amplification of the S1 and S4 genes and RFLP analysis.