RESUMEN
It is hypothesized that stem cells have the capability to form tumors after transplantation. Spermatogonial stem cells have proliferation potency and colonization ability related to express pluripotency genes such as c-Myc. The primary aim of this study is to investigate tumorigenicity ability of these cells after in vitro cultivation and inoculation in athymic animals. Spermatogonial stem cells from 3-5 day-old neonatal mice testes [NMRI] were cultured following two-step enzymatic digestion. After one month of culturing the spermatogonial stem cells, the obtained colonies were identified by Oct4 and PLZF markers. Expressions of Nanog, Oct4 and c-Myc pluripotency genes were subsequently studied. We subcutaneously inoculated 5 x 106 cells into athymic mice and assessed tumor formation after 8 weeks. Mouse embryonic stem cells [CCE line] were used as the positive control. Generated tumors were measured by a caliper. The colonies expressed Oct4 and PLZF proteins. Ratio of pluripotency gene expressions in these cells compared to embryonic stem cells significantly decreased [P = 0.05]. Mouse embryonic stem cells formed tumors however the spermatogonial colonies did not form any tumors. Mouse spermatogonial stem cells in comparison with embryonic stem cells are not capable of forming tumors in vivo. We have observed that the tumorigenic ability of these cells decreased significantly with down regulation of pluripotency gene expressions, particularly c-Myc. However, this study should be reassessed by using human tissue samples
RESUMEN
Endometrial integrin expression changes might be a reason for implantation failure in polycystic ovarian syndromes [PCOS]. Assessment of integrin genes and proteins expression upon endometrium in the PCOS experimental mouse model was the main goal of this study. 30 NMRI female mice were equally divided into control, experimental [PCOS; received estradiol valerate [40 mg/kg]] and sham group [received; olive oil]. After 8 weeks, each group was hyper stimulated by 7 IU PMSG and then, after 48hrs, 7 IU HCG was injected. Vaginal plaque was checked. After 5 days, Progesterone and estradiol levels and endometrial tissues were investigated to evaluate of alpha4, alphav, beta1 and beta3 integrins gene and protein by qPCR method and immunohistochemistry, respectively. Tissue samples were assessed and showed that level of progesterone was significantly decreased in PCOS group. Results of molecular part in the amount of alphav, beta3, beta1 and alpha4 gene expressions showed a great difference in beta3 and alphav genes expressions between experimental groups, alphav, beta3, alpha4 and beta1 proteins in the endometrial stroma in the control group were expressed, but they were not detected in PCOS group. According to the results, integrins had different expression patterns in different areas of the endometrium; such as epithelial and stromal. It seems that in PCOS, this pattern has changed and the results might have a great influence on implantation failure. Therefore, this study suggests that a great attention to this problem may be essential in patients who are involved
Asunto(s)
Animales de Laboratorio , Integrina alfa4 , Integrina alfa5 , Integrina beta3 , Integrina beta1 , Integrinas , Endometrio , Ratones , Expresión Génica , Implantación del EmbriónRESUMEN
It has been hypothesized that blastocyst integrin expression changes can affect the spontaneous miscarriage in polycystic ovarian syndromes [PCOS]. In this study, the profile of integrin genes and proteins was investigated on blastocyst of the PCOS experimental mouse model. 30 NMRI female mice were equally divided into 3 groups: control, experimental [PCOS that was injected estradiol valerate [40 mg/kg]]. After 8 weeks, each group was hyper stimulated by PMSG and HCG. Vaginal plaque was checked, and mice were investigated 5 days after the test. Progesterone and estradiol levels were determined; alpha4, alpha v, beta 1 and beta 3 integrin genes and protein of blastocysts were examined by real time PCR method and immunohistochemistry, respectively. Estradiol level was significantly increased [p = 0.035] in PCOS group. Based on our finding, the ratio of genes' expressions alphav, beta 3, beta 1 and alpha 4 in PCOS to control group was 0.479 +/- 0.01, 0.5 +/- 0.001, 2.7 +/- 0.4 and 1.023 +/- 0.2 respectively. Genes expression showed a great difference [p = 0.001] between beta 3, beta 1 and alphav in PCOS compared to other groups. alpha v and beta 3 integrin proteins expressed in all groups but intensity of these proteins in PCOS groups, was lower than other groups. Pattern of alpha v and beta3 integrins expression on the mouse blastocyst surface has an important effect during the implantation window. This pattern has changed in PCOS model and might have a great influence on implantation failure. Therefore, this experimental study suggests that a great attention to this problem may be essential in patients who are involved
Asunto(s)
Animales de Laboratorio , Integrina alfa4 , Integrina alfaV , Integrina beta3 , Integrina beta1 , Blastocisto , Ratones , Integrinas , Implantación del EmbriónRESUMEN
Sensory neurons in dorsal root ganglia [DRG] undergo apoptosis after peripheral nerve injury. The aim of this study was to investigate sensory neuron death and the mechanism involved in the death of these neurons in cultured DRG. In this experimental study, L5 DRG from adult mouse were dissected and incubated in culture medium for 24, 48, 72 and 96 hours. Freshly dissected and cultured DRG were then fixed and sectioned using a cryostat. Morphological and biochemical features of apoptosis were investigated using fluorescent staining [Propidium iodide and Hoechst 33342] and the terminal Deoxynucleotide transferase dUTP nick end labeling [TUNEL] method respectively. To study the role of caspases, general caspase inhibitor [Z-VAD.fmk, 100 microM] and immunohistochemistry for activated caspase-3 were used. After 24, 48, 72 and 96 hours in culture, sensory neurons not only displayed morphological features of apoptosis but also they appeared TUNEL positive. The application of Z-VAD.fmk inhibited apoptosis in these neurons over the same time period. In addition, intense activated caspase-3 immunoreactivity was found both in the cytoplasm and the nuclei of these neurons after 24 and 48 hours. Results of the present study show caspase-dependent apoptosis in the sensory neurons of cultured DRG from adult mouse