@article { author = {Ebrahimzadeh-Vesal, Reza and hokrgozar, Mohammad Ali and Nayernia, Karim and Teimoori-Toolabi, Ladan and Miryounesi, Mohammad and Nourashrafeddin, Seyedmehdi and Ranji, Najmeh and Modarressi, Mohammad Hosein}, title = {A vector-based system for the differentiation of mouse embryonic stem cells toward germ-line cells}, journal = {Iranian Journal of Basic Medical Sciences}, volume = {17}, number = {8}, pages = {566-570}, year = {2014}, publisher = {Mashhad University of Medical Sciences}, issn = {2008-3866}, eissn = {2008-3874}, doi = {10.22038/ijbms.2014.3190}, abstract = {Objective(s):To culture thein vitro mouse embryonic stem cells (mESCs) and to direct their  differentiation to germ-line cells; in present study we used a vector backbone containing the fusion construct Stra8-EGFP to select differentiated ES cells that entered meiosis.  Retinoic acid was used to differentiate embryonic stem cells to germ cells. Materials and Methods: A fragment of Stra8 gene promoter (-1400 to +7) was inserted in ScaI/HindIII multiple cloning site of pEGFP-1 vector. Theelectroporationwas done on embryonic stem cells and positive colonies were selected as puromycin-resistant after three weeks of treatment with puromycin. All-trans retinoic acid (RA) was used for differentiation of mESCs at final concentration of 10-5M. The expression of protamine 1 (Prm1) gene was checked as post meiotic marker in differentiated mESCs after 5, 10, 15, 21 and 30 days after RA induction. Results: The PCR amplification by specific primers for Stra8-EGFP fusion gene was detected in DNA sample from mESCs after electroporation and puromycin treatment.  GFP-positive mESC colonies were observed after 72 hr RA induction. The protamine 1 gene was expressed after 21 days of RA induction. Conclusion: In this study, we demonstrated the in vitro generation of mouse embryonic stem cells to germ cells by using a backbone vector containing the fusion gene Stra8-EGFP. The Stra8 gene is a retinoic acid-responsive protein and is able to regulate meiotic initiation.}, keywords = {Differentiation,Germ-line cells,Mouse embryonic stem cell,Vector-based system}, url = {https://ijbms.mums.ac.ir/article_3190.html}, eprint = {https://ijbms.mums.ac.ir/article_3190_ec4a8784d687d1bd9412eb77003aea81.pdf} }