BG02 cells were maintained in DC-HAIF media conditions for 32 passages. DC-HAIF consisted of DMEM/F12 (Invitrogen), 2% fatty acid-free Cohns fraction V BSA (Serologicals), 1x Nonessential amino acids, 50 U/ml Penicillin, 50 g/ml Streptomycin, 50 g/ml Ascorbic Acid, 10 g/ml bovine or human Transferrin, 0.1 mM ??ME (all from Invitrogen), 1x Trace Elements A, B & C (Mediatech), 10 ng/ml HRG1? (Peprotech), 10 ng/ml ActA (R&D Systems), 200 ng/ml LR3-IGF1 (JRH Biosciences), and 8 ng/ml FGF2 (Sigma or R&D Systems). HESCs were cultured in DC-HAIF on growth factor depleted Matrigel (BD Biosciences) diluted 1:200. Near-confluent hESC cultures were passaged by treating with 10 mg/ml (~2000 U/ml) Collagenase IV for three minutes, washing with 0.2% BSA in DMEM/F12, and gentle scraping to lift and break up the colonies. Colony pieces were centrifuged at 200g, aspirated, gently resuspended in DC-HAIF and typically split 1:3 to new plates.
Extracted molecule
total RNA
Extraction protocol
Trizol reagent (Invitrogen, Inc.)
Label
Biotin-Labeled cRNA
Label protocol
Illlumina RNA amplification kit, according to the manufacturers protocol
Hybridization protocol
Refer to the Illumina Gene Expression System Manual
Scan protocol
Refer to the Illumina Gene Expression System Manual
Description
Comparison of gene expression in human embyronic stem cells grown in MEF-CM or defined media conditions
Data processing
Raw data for each sample was output using the Illumina BeadStudio software
