|
| Status |
Public on Jan 02, 2014 |
| Title |
genomic DNA from primary osteocytes (donor 1) |
| Sample type |
genomic |
| |
|
| Source name |
NPT.O1
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
cell type: primary osteocytes (donor 1)
individual: donor 1
|
| Treatment protocol |
hASCs were differentiated to osteogenic and myogenic lineages by in vitro induction using specific culture medium. Osteogenesis was induced in the presence of DMEM with 10% FBS, dexamethasone (0.1 µM), ß-glycerophosphate (10 mM) and ascorbic acid-2-phosphate (50 µgr.ml-1). After 28 days, the culture medium was removed and cells were washed with PBS and processed. Myogenic induction of hASCs was obtained in the presence of DMEM with 10% FBS, hydrocortisone (50 μM) and horse serum 5% during 42 days.
|
| Growth protocol |
Human adipose-derived stem cells (hASCs) were established from adipose tissue of patients of 30-55 years of age. Lipoaspirates were digested with 0.075% collagenase Type I, prepared in PBS containing 1% BSA for 60 min at 37ºC with constant shaking, followed by filtration through a 100-µm filter. After washing with PBS, cells were treated with erythrocyte lysis buffer. Resultant cells were cultivated at 1,000 cells.cm-2 in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% of fetal bovine serum (FBS). Cultures were washed with buffer 24–48 hours after plating to remove unattached cells, and then re- fed with fresh medium. Only cells that were in passage 5 or 6 were used in these experiments. The human rhabdomyosarcoma (RD and TE.32.7) and osteosarcoma (MG-63) cell lines were obtained from the American Type Culture Collection (ATCC) (Rockland, MD, USA). Cell lines were maintained in monolayer cultures at 37ºC in an atmosphere containing 5% CO2, with DMEM supplemented with 10% FBS. Primary tissues from bone and muscle were obtained from healthy donors. All the samples were obtained after the donor had given their written informed consent, in accordance with the guidelines of the Ethics Committee of Bio-Health Research Foundation of Eastern Andalusia (Spain). DNA was extracted directly from fresh samples.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
DNA from all the samples was isolated by was applying the QIAamp® DNA Mini Kit (Qiagen Iberia, Spain) following supplier’s instructions.
|
| Label |
Cy5 and Cy3
|
| Label protocol |
Standard Illumina Protocol
|
| |
|
| Hybridization protocol |
bisulphite converted DNA was amplified, fragmented and hybridised to Illumina Infinium Human Methylation27 Beadchip using standard Illumina protocol
|
| Scan protocol |
Arrays were imaged using BeadArray Reader using standard recommended Illumina scanner setting
|
| Description |
primary osteocytes (donor 1)
|
| Data processing |
GenomeStudio software v2010,3
|
| |
|
| Submission date |
Nov 22, 2011 |
| Last update date |
Jan 02, 2014 |
| Contact name |
Antonio Gómez |
| E-mail(s) |
agomezm@idibell.cat
|
| URL |
http://www.pebc.cat/
|
| Organization name |
IDIBELL
|
| Department |
Epigenetics and Biology of Cancer Program
|
| Lab |
PEBC
|
| Street address |
Av. Gran Vía s/n km. 2,7.
|
| City |
L'Hospitalet de Llobregat |
| State/province |
Barcelona |
| ZIP/Postal code |
08907 |
| Country |
Spain |
| |
|
| Platform ID |
GPL8490 |
| Series (1) |
| GSE33896 |
Genome-wide CpG methylation during osteogenic and myogenic differentiation from adipose- derived stem cells. |
|
