|
| Status |
Public on Nov 10, 2007 |
| Title |
cumulus cells-late cleavage-rep6a |
| Sample type |
RNA |
| |
|
| Source name |
cumulus cells from oocyte resulting in a late cleavage embryo from patient 6
|
| Organism |
Homo sapiens |
| Characteristics |
The oocyte enclosed by these cumulus cells was fertilized normally (2PN) and developed into a 4-cell embryo with good morphology on day 2 and no multinucleated blastomeres present.
|
| Treatment protocol |
Cumulus cells were immediately lysed in 100 ml Trizol reagent (Invitrogen, Carlsbad, USA) supplemented with 1% (v/v) 2-mercapto-ethanol (Merck, Darmstadt, Germany), snap-frozen in liquid nitrogen and stored at -80°C (cumulus cells from one oocyte per vial).
|
| Growth protocol |
In consenting IVF patients, immediately following ultrasound-guided cumulus-oocyte-complex (COC) retrieval, a proportion of the cumulus cells surrounding a single oocyte were removed using a sharp needle.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, USA) according to the manufacturers’ instructions with some adaptations for the small quantity of RNA. RNA was precipitated with isopropyl alcohol for 2h and the RNA pellet was washed three times with 75% ethanol. To be able to track the small RNA pellet, 5 mg glycogen (Ambion, Woodward, USA) was added to the sample before RNA precipitation.
|
| Label |
biotin
|
| Label protocol |
Fifty ng total RNA was amplified using the two-cycle cDNA synthesis kit (Affymetrix, Santa Clara, USA) in combination with the MEGAscript T7 in vitro transcription system (Ambion) and biotin labeled with GeneChip IVT labeling kit (Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 11 ug of cRNA were hybridized for 16 hr at 45C on Human Genome U133 Plus 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip scanner 3000 / 7G.
|
| Description |
Comparison of cumulus cells from eight oocytes resulting in an early cleavage embryo (EC-CC; n=8) and from eight oocytes resulting in a non-EC embryo (NEC-CC; n=8).
|
| Data processing |
Affymetrix GCOS, TGT value: 500, Norm Factor: 1
|
| |
|
| Submission date |
Nov 06, 2007 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Aafke van Montfoort |
| E-mail(s) |
avmn@sgyn.azm.nl
|
| Organization name |
Academic hospital Maastricht
|
| Department |
Obstetrics & Gynaecology
|
| Lab |
IVF laboratory
|
| Street address |
P.O.Box 5800
|
| City |
maastricht |
| ZIP/Postal code |
6202 AZ |
| Country |
Netherlands |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE9526 |
Expression data from cumulus cells that surround oocytes resulting in early or late cleaving embryos |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE119087 |
