Establishing Clonal Cell Lines with Endothelial-Like Potential from CD9hi, SSEA-1− Cells in Embryonic Stem Cell-Derived Embryoid Bodies
Figure 2
Characterisation of E-RoSH cell lines.
a) Cellular morphology of E-RoSH2.1 and RoSH2 cells in sub-confluent cultures;
b,c) Pairwise comparison of global gene expression between E14 ESCs and E-RoSH2.1/E-RoSH3.2, and RoSH2 and E-RoSH2.1/E-RoSH3.2.
Global gene expression analysis of E14 ESCs, E-RoSH, RoSH were performed by hybridizing total RNA from two biological samples each of E14 ESCs, E-RoSH2.1, E-RoSH3.2, and RoSH2 with Illumina BeadArray containing about 24,000 unique features;
d) Biological processes in which 1115 highly expressed genes in ESCs have statistically significant higher frequencies (p<0.05).
The biological processes were (left to right): neuron differentiation, neuron development, nervous system development, neuron maturation, nerve ensheathment, cellular nerve ensheathment, ionic insulation of neurons by glial cells, myelination, transmission of nerve impulse regulation of action potential, neurophysiological process, neuron morphogenesis during differentiation, neurite morphogenesis, axonogenesis, cell development, development, system development, cell maturation, cell differentiation, cellular morphogenesis, regulation of gene expression, epigenetic Imprinting, gametogenesis, morphogenesis, sexual reproduction, cell-cell signalling, cell communication, metabolism, cell organization and biogenesis, regulation of biological process, macromolecule catabolism, carbohydrate catabolism, cellular carbohydrate catabolism, monosaccharide catabolism, hexose catabolism, glucose catabolism, glycolysis, cellular macromolecule catabolism, cellular catabolism, alcohol catabolism, carbohydrate metabolism, cellular carbohydrate metabolism, monosaccharide metabolism, hexose metabolism, glucose metabolism, main pathways of carbohydrate metabolism alcohol metabolism, generation of precursor metabolites and energy, energy derivation by oxidation of organic compounds;
e) Biological processes in which 1263 highly expressed genes in ESCs have statistically significant higher frequencies (p<0.05).
The biological processes were (left to right): proteolysis, protein metabolism, cellular protein metabolism, cellular macromolecule metabolism, macromolecule metabolism, memory and vasculature development;
f,g, h, i) Relative gene expression analysis by quantitative RT-PCR analysis.
The expression level was normalized against that of ESCs and expressed as a logarithmic function;
j) Western blot analysis for pluripotency-associated gene products in cell extracts of ESCs, EBs and E-RoSH2.1 cells.