Study on immortal conditions of chicken embryonic stem cells

Abstract

In recent years, considerable attention has been paid to chicken embryonic stem cells (ESCs) studies in relation to extensive applications in gene therapy and regenerative medicine. However, the approaches used are still immature. In this study, we showed that the chicken ESCs clones with a clear border can express alkaline phosphatase and marker proteins such as SSEA‐1, SOX2, and OCT4 stably. In addition, culture medium containing 10 μmol/L of vitamin C (VC) could significantly promote the proliferation of ESCs cells. Moreover, ESCs transfected with p:enhanced green fluorescent protein (pEGFP)‐hTERT could be subcultured more than tenth generations in culture medium containing exogenous factors (mLIF + bFGF + hSCF) and VC, and these ESCs clone could still be regenerated following cryopreservation. Quantitative real‐time polymerase chain reaction results showed that there was no significant difference between SSEA‐1, SOX2, and OCT4 expression during ESCs immortalization and that the tenth generation of ESCs was still able to express marker proteins SSEA‐1, SOX2, and OCT4. Our results showed that an immobilized system for ESCs was established, and the ESCs were cultured in vitro maintaining their pluripotency.

Transcriptome Sequencing and Comparative Analysis of Amphoteric ESCs and PGCs in Chicken (Gallus gallus)

“…For decades, chickens have played a vital role in animal research as alternatives and outbred experimental species to humans to compensate for ethical constraints [2][3][4]. It works especially well when isolating and gaining a considerable number of pluripotent embryonic stem cells (ESCs) and primordial germ cells (PGCs) is much easier and convenient in chickens than other species due to the in ovo embryonic development [5,6]. This unique advantage accelerates the application of chicken ESCs and PGCs in studying the mechanism of human germ cell development and differentiation [7,8].…”

“…embryonic stem cells; 2 primordial germ cells;3 the ratio of absorbance at 260 and 280 nm;4 the ratio of absorbance at 260 and 230 nm;5 the ratio of 28S/18S ribosomal RNA; 6 RNA integrity number.…”