Global gene expression profiling reveals similarities and differences among mouse pluripotent stem cells

ABSTRACT: We present a compendium DNA microarray analysis of multiple mouse ESCs and EGCs from different genetic backgrounds (strains 129 and C57BL/6) cultured under standard conditions and in differentiation-promoting conditions by the withdrawal of Leukemia Inhibitory Factor (LIF) or treatment with retinoic acid (RA). All pluripotent cell lines showed similar gene expression patterns, which separated them clearly from other tissue stem cells with lower developmental potency. Differences between pluripotent lines derived from different sources (ESC vs. EGC) were smaller than differences between lines derived from different mouse strains (129 vs. C57BL/6). Multiple ESC and EGC cell lines were cultured without feeders on gelatin coated, 6-well plates, 100,000 cells/well (10,000 cells/cm2) in 3 conditions: 1) in complete ES medium: DMEM, 15% FBS; LIF (ESGRO) 1000 u/ml; 1mM sodium pyruvate; 0.1 mM NEAA, 2 mM glutamate, 0.1 mM beta-mercapto ethanol, and penicillin/streptomycin (50U/50ug per ml); 2) in ES medium without LIF; 3) in complete ES medium plus 1 uM RA. Cells were incubated at 37 0C; 5% CO2. Medium was changed daily. On the day 3 Trizol was added, RNA extracted using Phase lock gelTM (Eppendorf) columns according to manufacturer protocol. RNA was precipitated with isopropanol, washed with 70% ethanol and dissolved in DEPC dH2O.

ORGANISM(S): Mus musculus

SUBMITTER: Nabeebi Shaik 

PROVIDER: E-GEOD-5914 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress