|
| Status |
Public on Jul 21, 2011 |
| Title |
Human pancreatic beta cell-enriched cell, biological replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
Pool of pancreatic beta cell-enriched fractions, obtained from 3 non-selected adult human pancreases (Beta Cell Bank at Diabetes Research Center, Brussels, Belgium)
|
| Organism |
Homo sapiens |
| Characteristics |
tissue source: 3 non-selected adult human pancreases
pancreatic cell type: adult pancreatic endocrine cells
|
| Treatment protocol |
Suspensions of human beta and duct cells were maintained 2-3w in culture as described ( Diabetes 2001,49: 571-579 ) with no specific treatment.
|
| Growth protocol |
We hybridized 3 different pools of 6 µg RNA obtained from 10 human organs in total: pool 1 (45% insulin+, from n=3 donor organs), pool 2 (65% ins+, from n=3 donor organs) and pool 3 (55% insulin+, from n=4 donor organs). Average donor age and BMI were 47±10 yrs and 26±4, respectively. Pancreatic duct cells-enriched preparations contained 85±7% cytokeratin 19+ cells with 4±1% insulin+ cells and 6±4% glucagon+ cells and were isolated as described by Heimberg H. et al.( Diabetes 2001,49: 571-579 ).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
After fragmentation at 94°C for 35 min in fragmentation buffer (40mM Tris, 30mM magnesium acetate, 10mM potassium acetate), the labelled cRNA was hybridized for 16h to the Affymetrix HG133A
|
| Scan protocol |
Arrays were stained with phycoerythrin-streptavidin with the Affymetrix Fluidics Station 400, and scanned in the Affymetrix GeneArray2500scanner.
|
| Description |
Beta cell-a1
each biological replicate obtained from pooled cultures of 3-4 human organs, 10 donor organs in total
Beta Cell Bank at Diabetes Research Center, Brussels, Belgium
|
| Data processing |
Scanned arrays were analyzed with dChip model-based expression analysis to the array with the median intensity. Using the PM-MM model with mismatch correction, a 90% confidence interval was calculated for the fold change in gene expression in each comparison, and the lower limit of this interval (LCB) was used as a measure of differential gene expression.
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| |
|
| Submission date |
Jul 19, 2011 |
| Last update date |
Aug 05, 2011 |
| Contact name |
Geert A. Martens |
| E-mail(s) |
[email protected]
|
| Organization name |
Vrije Universiteit Brussel
|
| Department |
Diabetes Research Center
|
| Street address |
Laarbeeklaan 103
|
| City |
Brussels |
| ZIP/Postal code |
1090 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE30803 |
Genome-wide mRNA profiling of adult human pancreatic beta and duct cells in comparison to other human tissues |
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