Abstract
Pluripotent Embryonic Stem cell (ESC) lines can be derived from a variety of sources. Mouse lines derived from the early blastocyst and from primordial germ cells (PGCs) can contribute to all somatic lineages and to the germ line, whereas cells from slightly later embryos (EpiSC) no longer contribute to the germ line. In chick, pluripotent ESCs can be obtained from PGCs and from early blastoderms. Established PGC lines and freshly isolated blastodermal cells (cBC) can contribute to both germinal and somatic lineages but established lines from the former (cESC) can only produce somatic cell types. For this reason, cESCs are often considered to be equivalent to mouse EpiSC. To define these cell types more rigorously, we have performed comparative microarray analysis to describe a transcriptomic profile specific for each cell type. This is validated by real time RT-PCR and in situ hybridisation. We find that both cES and cBC cells express classic pluripotency-related genes (including cPOUV/OCT4, NANOG, SOX2/3, KLF2 and SALL4), whereas expression of DAZL, DND1, DDX4 and PIWIL1 defines a molecular signature for germ cells. Surprisingly, contrary to the prevailing view, our results also suggest that cES cells resemble mouse ES cells more closely than mouse EpiSC.
Highlights
Embryonic stem (ES) cells were first generated from mouse embryos in 1981 (Evans and Kaufman, 1981; Martin, 1981), in the primates (Thomson et al, 1995) including human (Thomson et al, 1998)
A pluripotent cell is defined as having the ability to contribute to many, or even all, cell types
Pluripotency can be demonstrated in vivo, when cells are introduced into a recipient embryo, and in vitro, when cells differentiate into derivatives of the three primary embryonic cell layers
Summary
Embryonic stem (ES) cells were first generated from mouse embryos in 1981 (Evans and Kaufman, 1981; Martin, 1981), in the primates (Thomson et al, 1995) including human (Thomson et al, 1998). Functional PGCs can be obtained from the embryonic blood of stage 14-17 HH (Hamburger and Hamilton, 1951) embryos (Naito et al, 2004; van de Lavoir et al, 2006a,b; Macdonald et al, 2010, 2012; Park and Han, 2012) or from the gonads of stage 28-30 (Hamburger and Hamilton, 1951) embryos (Ha et al, 2002; Park et al, 2003; Song et al, 2014) These PGCs can be established and maintained in culture using a similar medium as described for cES (Pain et al, 1996), but supplemented with higher concentrations of FGF and SCF and by promoting the non-adherent floating cells that emerge in culture (van de Lavoir et al, 2006a,b; Macdonald et al, 2010). These cells appear very promising for generating genetically modified chickens (Park and Han, 2012; Macdonald et al, 2012; Schusser et al, 2013)
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