Abstract
Primordial germ cells (PGCs) are the precursors of adult germ cells, and among the embryonic stem-like cells in the bird embryo, only they can transmit the genetic information to the next generation. Despite the wide range of applications, very little is known about the mechanism that governs primordial germ cell self-renewal and differentiation. As a first step, we compared 12 newly established chicken PGC lines derived from two different chicken breeds, performing CCK-8 proliferation assay. All of the lines were derived from individual embryos. A significant difference was found among the lines. As microRNAs have been proved to play a key role in the maintenance of pluripotency and the cell cycle regulation of stem cells, we continued with a complex miRNA analysis. We could discover miRNAs expressing differently in PGC lines with high proliferation rate, compared to PGC lines with low proliferation rate. We found that gga-miR-2127 expresses differently in female and male cell lines. The microarray analysis also revealed high expression level of the gga-miR-302b-3p strand (member of the miR-302/367 cluster) in slowly proliferating PGC lines compared to the gga-miR-302b-5p strand. We confirmed that the inhibition of miR-302b-5p significantly increases the doubling time of the examined PGC lines. In conclusion, we found that gga-miR-181-5p, gga-miR-2127, and members of the gga-miR-302/367 cluster have a dominant role in the regulation of avian primordial germ cell proliferation.
Highlights
The avian embryo provides an excellent model system for developmental and cell biology
We are presenting a detailed study on the proliferation and developmental properties of 12 chicken Primordial germ cells (PGCs) lines
Among the 21 PC lines, only 4 (19%), in case of the 10 GFP lines, only 2 (20%) female PGC lines were detected by using sex PCR (Figure 1)
Summary
The avian embryo provides an excellent model system for developmental and cell biology. It has been widely used in early embryogenesis, toxicology, and stem cell studies [1,2,3]. Primordial germ cells (PGCs) are the precursors of adult germ cells and have remarkable potential in developmental biology-, disease modelling-, and drug development-related research applications [4]. They are accessible from the embryonic blood and are suitable for genome preservation (gene banks) [5]. They tend to regulate and express in multiple biological processes such as cell growth, differentiation, apoptosis, and development [8]
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