Study on immortal conditions of chicken embryonic stem cells

This study not only optimized the isolation and culture of chicken embryonic stem cells (ESCs) but also investigated the immortalization of chicken ESCs by transfecting chicken ESCs with pEGFP ‐hTERT and culturing transfected cells in conditioned medium with exogenous factors and 10 μmol/L vitamin C. The results showed that cells can passage to tenth generations. Following indirect immunofluorescence and quantitative real‐time polymerase chain reaction detection indicated that the cells still have pluripotency. The method of efficient separation of chicken ESCs and the immobilized system was established, which provides the basis for unlimited chicken ESCs proliferation. AbstractIn 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 followin...
Source: Journal of Cellular Biochemistry - Category: Biochemistry Authors: Tags: RESEARCH ARTICLE Source Type: research