|
| Status |
Public on Jan 11, 2015 |
| Title |
AdiposeDerivedStemCells7_osteo_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Adipose-derived stem cells, donor 7, passage 3, osteogenic differentiation
|
| Organism |
Homo sapiens |
| Characteristics |
differentiation status: osteogenic differentiation
original cell type: Adipose-derived stem cells
|
| Treatment protocol |
Cells were treated with osteogenic-induction medium, prepared from DMEM-high glucose plus osteogenic stimulatory supplement, dexamethasone (final concentration 10^-8 M) and ascorbic acid (50 μg/ml). When formation of cell multilayers became evident, β-glycerophosphate was added to the osteogenic-induction medium (10^-6 M) and differentiation was performed for another 2 weeks. Control cells were simultaneously cultured in the complete proliferation medium.
|
| Growth protocol |
Sections of adipose tissue were digested with 0.2% collagenase type I in DMEM at 37°C and filtered through 100 μm filter. Cells were grown in DMEM, supplemented with 10% FCS and antibiotics, in a humidified atmosphere with 5% CO2 at 37°C and passaged when reached subconfluence of 75-90%.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from 2 mln. cells on average with mirVana miRNA Isolation Kit according to manufacturer’s protocol (Life Technologies). Extracted RNA was quantified using NanoDrop-2000 spectrophotometer (Thermo Fisher Scientific) and quality was monitored with Agilent 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 labeled cRNA was prepared from 200 ng of purified RNA using Low Input Quick Amp Labeling Kit, One Color and RNA Spike-In Kit, One Color acording to manufacturer's protocol (Agilent Technologies), followed by purification with RNeasy Mini Kit (Qiagen). Dye incorporation and cRNA yield was measured with NanoDrop 2000.
|
| |
|
| Hybridization protocol |
All amount of labeled cRNA was prepared for hybridization using Gene Expression Hybridization Kit following manufacturer's protocol (Agilent technologies). Samples were hybridized onto Human Gene Expression (v2) 8×60K microarrays for 17 hours at 65°C in a rotating hybridization oven (Agilent Technologies).
|
| Scan protocol |
Microarrays were scanned immediately after washing using SureScan microarray scanner (Agilent Technologies).
|
| Description |
Gene expression in osteogenically differentiated ADSCs from donor 7
|
| Data processing |
Scanned images were analyzed with Feature Extraction software v10.7 (Agilent Technologies) using default parameters (protocol GE1_107_Sep09, grid 039494_D_F_20120628 for ADSC4 and 039494_D_F_20140228 for ADSC6 and ADSC7). Background subtracted and spatially detrended signals were obtained.
|
| |
|
| Submission date |
Dec 01, 2014 |
| Last update date |
Jan 11, 2015 |
| Contact name |
Kristina Daniunaite |
| Organization name |
Vilnius University
|
| Department |
Institute of Biosciences
|
| Lab |
Human Genome Research Centre
|
| Street address |
Sauletekio Ave. 7
|
| City |
Vilnius |
| ZIP/Postal code |
LT-10257 |
| Country |
Lithuania |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE63754 |
Gene expression profile of osteogenically differentiated human adipose-derived stem cells |
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