animal
Masoumeh Zareh; Seyed Ziaedin Mirhoseini; Seyedeh Nafiseh Hasani; Shahrokh Ghovvati
Abstract
Introduction: The main challenge of primordial germ cells (PGC) is their lack of proliferation and self-renewal in the culture medium. One method for inducing the pluripotency of PGC cells is to manipulate intracellular signaling pathways such as TGF-β, and the use of growth factors and small molecules ...
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Introduction: The main challenge of primordial germ cells (PGC) is their lack of proliferation and self-renewal in the culture medium. One method for inducing the pluripotency of PGC cells is to manipulate intracellular signaling pathways such as TGF-β, and the use of growth factors and small molecules is one of the ways to achieve this goal. Materials and methods: Chicken gonadal PGC cells were cultured and incubated with a concentration of 50,000 cells per well in a 24-well plate coated with Matrigel. The experimental groups included four groups: control (basic medium for PGCs culture), treatment with small molecule IDE1 (100 nM; Stemgent, USA, 04-0026), treatment with growth factor A Activin (25 ng/ml; R&D Systems, 338-AC) and treatment with SB431542 (10 µM; Cayman Chemical, 13031) with three replicates from each group. In order to check the amount of cell proliferation, PGC cells were counted in time intervals of 7, 14, 21 days after the treatment with a hemocytometer. The activity of TGF/ẞ signaling pathway was evaluated by examining the expression of SMAD2, SMAD3 and LFTTY1 genes by qRT-PCR method. Results: The effect of Activin A and IDE1 led to an increase in the proliferation of PGCs cells to more than 4 times compared to the control group, and in contrast to the SB431542 group, it led to a decrease in cell proliferation.
