Secondary SKPs spheres were dissociated and cultured at 50,000 cells/ml in SKPs basal growth medium (DMEM-F12, 3:1 and 40 ng/ml FGF2, 20 ng/ml EGF, NSCP were cultured in growth medium containing 1% P/S, 2% N2 supplement , 25 ng/ml neuregulin-1β, and 5 μM forskolin on coated plates (4 μg/ml laminin, 30 μg/ml poly-D-lysine hydrobromide)
Growth protocol
Cells were cultured at 50,000 cells/ml in SKPs basal growth medium (DMEM-F12, 3:1 and 40 ng/ml FGF2, 20 ng/ml EGF, NSCP were cultured in growth medium containing 1% P/S, 2% N2 supplement , 25 ng/ml neuregulin-1β, and 5 μM forskolin on coated plates (4 μg/ml laminin, 30 μg/ml poly-D-lysine hydrobromide) 2% B27, and 1 μg/ml fungizone) and allowed to form spheres
Extracted molecule
total RNA
Extraction protocol
RNA was extracted using the Qiagen Rneasy kit according to the manufacturer's instructions
Label
biotin
Label protocol
cDNA was prepared using the Ambion WT Expression Kit using 400 ng of total RNA
Hybridization protocol
Affymetrix Rat Gene 2.0 ST arrays were washed, stained and scanned according to the Fluidics Protocol FS450_0002 using 5.5 ug of cDNA
Scan protocol
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000
Description
Gene expression of neonatal rat NSCPs
Data processing
The data were analyzed using the oligo package of Bioconductor in R. The data were background corrected, normalized with quantile normalization, transformed into the log2 scale, and summarized into probesets using the Robust Multichip Analysis (RMA) method.
