Abstract
Chicken embryonic stem cells (cESCs) obtained from stage X embryos provide a novel model for the study of avian embryonic development. A new way to maintain cESCs for a long period in vitro still remains unexplored. We found that the cESCs showed stem cell-like properties in vitro for a long term with the support of DF-1 feeder and basic culture medium supplemented with human basic fibroblast growth factor (hbFGF), mouse stem cell factor (mSCF), and human leukemia inhibitory factor (hLIF). During the long culture period, the cESCs showed typical ES cell morphology and expressed primitive stem cell markers with a relatively stable proliferation rate and high telomerase activity. These cells also exhibited the capability to differentiate into cardiac myocytes, smooth muscle cells, neural cells, osteoblast, and adipocyte in vitro. Chimera chickens were produced by cESCs cultured for 25 passages with this new culture system. The experiments showed that DF-1 was the optimal feeder and hbFGF was an important factor for maintaining the pluripotency of cESCs in vitro.
Highlights
Since embryonic stem cells (ESCs) were first obtained from mouse embryos [1], tremendous efforts have been made to study ESCs varying from species, like human [2], zebrafish, porcine [3], and bovine [4]
The results revealed that the Chicken embryonic stem cells (cESCs) expressed FGFRs (Figure S2) and the human basic fibroblast growth factor (hbFGF) significantly increased the area of colonies, compared to human leukemia inhibitory factor (hLIF) and mouse stem cell factor (mSCF) (Figure 1(b))
Our results suggested that this newly established culture system is able to maintain the blastodermal cells with stemlike features over 48 passages in vitro
Summary
Since embryonic stem cells (ESCs) were first obtained from mouse embryos [1], tremendous efforts have been made to study ESCs varying from species, like human [2], zebrafish, porcine [3], and bovine [4]. The pluripotency of a chicken blastoderm at stage X was confirmed by chimera production and differentiation into various lineages [5, 6]. Pain and his colleagues maintained the cESCs (chicken embryonic stem cells) in vitro for a long time leading to the establishment of the cell line 9N2-5 [7, 8].
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