Moreover, ES cell differentiation in vitro recapitulates events that take place
during early
embryonic development including the
formation of the three germ layers of ectoderm, mesoderm and endoderm, and the emergence of endothelial, hematopoietic, cardiac, neuronal and hepatic or pancreatic cells [8], [9].
For example, clusters containing genes that are upregulated
during the course of ES cell differentiation (Table 3) include in order of time of expression: cluster 30 that represents genes which take part in the
formation of the three
embryonic germ layers
during gastrulation, i.e., Goosecoid, Cerberus like 1 homolog, Wnt3, Mesp1, Mixl1, mEomes and Even - skipped 1; cluster 15 containing molecular regulators of early mesoderm
development including Bmp2, Bmp5, Msx1, Msx2, Cripto, Tbx20, Hey2, Smad6, Vegfr2 (Kdr), Foxf1 and Hand1; cluster 20, which comprises regulatory and structural genes linked to hemopoiesis such as Gata1, Nfe2, Klf1, Tie1, hemoglobins (Hba - x, Hbb - b1) and Glycophorin A; cluster 12, which is rich in genes involved in cardiac
development, e.g., Mef2c, Myl4, cardiac Troponin T2, Tropomodulin 1, myosin binding protein C, Bves, Angiopoietin 1 and Angiopoietin 2; and, cluster 4, which consists mostly of genes associated with neuronal
development and differentiation, for example, Neurog1, Neurog2, Olig2, Nkx6.1, Neurod4, Pou3f2, Pou3f4, Cacna2d3, Cacng4, Kcnq2 and EphA5.
Taking into account that ES cells are isolated at
embryonic day 3.5 post fertilization, the sequential appearance of genes specific for gastrulation, mesoderm
formation, hemopoiesis, cardiopoiesis and neurogenesis
during ES cell differentiation follows the timing of comparable developmental stages in
embryonic development.