원문정보
초록
영어
Chicken embryonic stem (ES) cells have great potential and provide a powerful tool to investigate embryonic development and to manipulate genetic modification in a genome. However, very limited studies are available on the functional characterization and robust expansion of chicken ES cells compared to other species. Here, we have developed a method to generate chicken embryonic stem cell-like cells under pluripotent culture conditions. The chicken embryonic stem cell-like cells were cultivated long-term over several passages of culture without loss of pluripotency in vitro and had the specific expression of key stem cell markers. Furthermore, they showed severe changes in morphology and a significant reduction in pluripotent genes after siRNA-mediated NANOG knockdown. Collectively, these results demonstrate the efficient generation of chicken embryonic stem cell-like cells from EGK stage X blastoderm-derived singularized cells and will facilitate their potential use for various purposes, such as biobanking genetic materials and understanding stemness in the fields of animal biotechnology.
목차
INTRODUCTION
MATERIALS AND METHODS
Isolation and cultivation of EGK stage X blastoderm-derivedsingularized cells
Preparation of CF1 mouse embryonic fibroblasts(MEFs) as a feeder layer
siRNA-mediated knockdown of the chicken NANOG gene in vitro
Periodic acid-Schiff (PAS) and alkaline phosphatase (AP) staining
Quantitative RT‒PCR
Statistical analysis
RESULTS
Generation and characterization of chicken embryonic stem cell-like cells
Effect of siRNA-mediated knockdown of chicken NANOG
DISCUSSION
CONCLUSION
REFERENCES