In vitro improvement of quail primordial germ cell expansion through activation of TGF‐beta signaling pathway. Issue 6 (7th March 2018)

• Record Type: Journal Article
• Authors: Yakhkeshi, Saeed; Rahimi, Shaban; Sharafi, Mohsen; Hassani, Seyedeh‐Nafiseh; Taleahmad, Sara; Shahverdi, Abdolhossein; Baharvand, Hossein
• Is Part Of: Journal of cellular biochemistry. Volume 119:Issue 6(2018)
• Page Start: 4309
• Page End: 4319
• Publication Date: 2018-03-07
• Subjects: IDE1 small molecule -- primordial germ cell -- signaling pathway -- quail embryo; Cytochemistry -- Periodicals; 572
• DOI: 10.1002/jcb.26618
• Languages: English
• ISSNs: 0730-2312

Abstract

Abstract: Avian primordial germ cells (PGCs) have valuable potentials to cell‐based approaches for transgenic bird production. In this regard, improvement of avian PGC expansion in vitro is necessary. Among experimental avian species, quail is a good model for transgenic technology, especially due to its short generation time. In the present study, we have examined the proliferative effects of transforming growth factor β (TGF‐β) on the quail PGCs. After isolation of quail PGCs from blood (Hamburger‐Hamilton [HH stages 13‐15]) and gonads (HH stages 28‐30), these cells were cultured on quail embryonic fibroblasts (QEF). Our results indicated th at cultured gonadal‐derived PGCs proliferated 400 times in comparison to 100 times for blood PGCs over 40‐50 days. Upon in vitro exposure to TGF‐β inducers by Activin or the inducer of definitive endoderm 1 (IDE1) small molecule, the number of gonad PGCs significantly increased to 26% and 64%, respectively. In contrast, inhibition of the TGF‐β signaling pathway by SB431542 resulted in a significant reduction in the numbers of PGCs ( P < 0.001). Moreover, Phosphorylation of SMAD2/3 in the IDE1 group was higher compared to the Activin‐treated ones. We confirmed the PGC identification with periodic acid‐Schiff (PAS) staining, anti‐SSEA1, β‐catenin, β‐integrin, and Nanog immunofluorescence staining. Exogenously IDE1 treated‐PGCs migrated toward the embryonic gonads after transplantation into the heart of the recipient embryo at HH stages 13‐15. Our results suggested that the application of IDE1 small molecule into the culture of quail PGCs represented a step toward achieving efficient expansion of the avian PGCs. Abstract : TGF‐β inducers by Activin or IDE1 increased the number of gonadal PGCs significantly. IDE1 increased the number of PGCs more than Activin. Phosphorylation of SMAD2/3 in the IDE1 group was higher than the Activin‐treated ones.