PMID- 35035539
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220118
IS  - 1984-3143 (Electronic)
IS  - 1806-9614 (Print)
IS  - 1806-9614 (Linking)
VI  - 18
IP  - 4
DP  - 2021
TI  - The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine 
      spermatogonial stem cells.
PG  - e20200257
LID - 10.1590/1984-3143-AR2020-0257 [doi]
LID - e20200257
AB  - 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 microM), 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 microM 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 microM 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 microM in the Y-27632 
      dose. In the 20 microM 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 microM 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 microM in 
      comparison with the control group. In conclusion, the present study demonstrated 
      that Y-27632 at a concentration of 5-10 microM provided optimal culture conditions 
      for the primary culture of ovine SSCs.
FAU - Emamdoust, Fatemeh
AU  - Emamdoust F
AUID- ORCID: 0000-0002-3088-093X
AD  - Department of Animal Science Faculty of Agriculture and Natural Resources, 
      Science and Research Branch Islamic Azad University Tehran Iran Department of 
      Animal Science, Faculty of Agriculture and Natural Resources, Science and 
      Research Branch, Islamic Azad University, Tehran, Iran.
FAU - Aminafshar, Mehdi
AU  - Aminafshar M
AUID- ORCID: 0000-0002-6238-0497
AD  - Department of Animal Science Faculty of Agriculture and Natural Resources, 
      Science and Research Branch Islamic Azad University Tehran Iran Department of 
      Animal Science, Faculty of Agriculture and Natural Resources, Science and 
      Research Branch, Islamic Azad University, Tehran, Iran.
FAU - Zandi, Mohammad
AU  - Zandi M
AUID- ORCID: 0000-0002-0523-1719
AD  - Department of Agriculture Iranian Research Organization for Science and 
      Technology Tehran Iran Department of Agriculture, Iranian Research Organization 
      for Science and Technology, Tehran, Iran.
FAU - Sanjabi, Mohammad Reza
AU  - Sanjabi MR
AUID- ORCID: 0000-0003-4202-2892
AD  - Department of Agriculture Iranian Research Organization for Science and 
      Technology Tehran Iran Department of Agriculture, Iranian Research Organization 
      for Science and Technology, Tehran, Iran.
LA  - eng
PT  - Journal Article
DEP - 20220107
PL  - Brazil
TA  - Anim Reprod
JT  - Animal reproduction
JID - 101272804
PMC - PMC8747935
OTO - NOTNLM
OT  - Y-27632
OT  - primary culture
OT  - sheep
OT  - spermatogonial stem cells
COIS- Conflict of Interest: The authors have no conflict of interest to declare.
EDAT- 2022/01/18 06:00
MHDA- 2022/01/18 06:01
PMCR- 2022/01/07
CRDT- 2022/01/17 05:55
PHST- 2021/01/07 00:00 [received]
PHST- 2021/11/19 00:00 [accepted]
PHST- 2022/01/17 05:55 [entrez]
PHST- 2022/01/18 06:00 [pubmed]
PHST- 2022/01/18 06:01 [medline]
PHST- 2022/01/07 00:00 [pmc-release]
AID - arAO20200257 [pii]
AID - 10.1590/1984-3143-AR2020-0257 [doi]
PST - epublish
SO  - Anim Reprod. 2022 Jan 7;18(4):e20200257. doi: 10.1590/1984-3143-AR2020-0257. 
      eCollection 2021.