التفاصيل البيبلوغرافية
| العنوان: |
The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells |
| المؤلفون: |
Emamdoust, Fatemeh, Aminafshar, Mehdi, Zandi, Mohammad, Sanjabi, Mohammad Reza |
| المصدر: |
Animal Reproduction. January 2021 18(4) |
| بيانات النشر: |
Colégio Brasileiro de Reprodução Animal, 2021. |
| سنة النشر: |
2021 |
| مصطلحات موضوعية: |
spermatogonial stem cells, primary culture, Y-27632, sheep |
| الوصف: |
The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-old’s lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V–FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days’ culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs. |
| نوع الوثيقة: |
article |
| وصف الملف: |
text/html |
| اللغة: |
English |
| تدمد: |
1984-3143 |
| DOI: |
10.1590/1984-3143-ar2020-0257 |
| URL الوصول: |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-31432021000400205 |
| Rights: |
info:eu-repo/semantics/openAccess |
| رقم الانضمام: |
edssci.S1984.31432021000400205 |
| قاعدة البيانات: |
SciELO |
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