Abstract
The ES cell can provide a useful system for studying differentiation and development in vitro and a powerful tool for producing transgenic animalds. To investigate the culture condition of chicken embryonic stem (CES) cells which can retain their multipotentiality or totipotency, three kinds of feeder layer cells, SNL cells, primary mice embryonic fibroblasts (PMEF) cells and primary chicken embryonic fibroblasts (PCEF) cells, were used as the feeder cells in media of DMEM supplemented with leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and stem cell factor (SCF) for co-culture with blastoderm cells from stage X embryos of chicken. The alkaline phosphatase (AKP) test, differentiation experiment in vitro and chimeric chicken production were carried out. The results showed that culture on feeder layer of PMEF yielded high quality CES cell colonies. The typical CES cells clone shape revealed as follows: nested aggregation (clone) with clear edge and round surface as well as close arrangement within the clone. Strong alkaline phosphatase (AKP) reactive cells were observed in the fourth passage cells. On the other hand, the fourth passage CES cells could differentiate into various cells in the absence of feeder layer cells and LIF in vitro. The third and fourth passage cells were injected into the subgerminal cavity of recipient embryos at stage X. Of 269 Hailan embryos injected with CES cells of Shouguang Chickens, 8.2% (22/269) survived to hatching, 5 feather chimeras had been produced. This suggests that an effective culture system established in this study can promote the growth of CES cells and maintain them in the state of undifferentiated and development, which lays a solid foundation for the application of CES cells and may provide an alternative tool for genetic modification of chickens.
Highlights
Embryonic stem (ES) cells which are derived from the blastocyst stage of the embryo have the same characteristics as embryo cells They are capable of differentiating into all types of cells and can be isolated and cultured for long-term in vitro
The aim of the present study was to compare the effect of three feeder layer cells, three cytokines and other factors on the growth of chicken ES cells in order to set up the optimum culture condition allowing the growth and characterization of chicken ES cells
Growth was started from chicken blastoderms collected at stage X (Eyal-Giladi and Kochar, 1976) that were known to contain cells that contribute to the germ line in chimeras (Carsience et al, 1993; Petitte et al, 1990) in an attempt to develop cultures of chicken totipotent embryonic cells, After cultured for 24 h the CB cellcolonies composed of 10-20 cells appeared on the feeder layer cells and these colonies became more obvious after another 24 h. 3-4 days later we could see some nest-like, Alkaline phosphatase reaction hill-like, sunflower-like colonies and 5 days later these
Summary
Embryonic stem (ES) cells which are derived from the blastocyst stage of the embryo have the same characteristics as embryo cells They are capable of differentiating into all types of cells and can be isolated and cultured for long-term in vitro. These cultured cells can participate in the development of all cell lineages including the germ line when they are implanted into host blastocysts. Etches et al (1997); Du Lixin (2002) had maintained these blastodermal cells in vitro for 48h and Chinese Academy of Agricultural Sciences Beijing 100094, Received November 14, 2002; Accepted April 4, 2003 proven that genetically modified avian ES cells continued to have the ability to join in the somatic and the germ line
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
