|
| Status |
Public on Oct 02, 2012 |
| Title |
Fibroblast mEF1 |
| Sample type |
RNA |
| |
|
| Source name |
Fibroblast
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Fibroblast
genetic background: C57BL/6
|
| Treatment protocol |
HB isolated from a pool of fetal (ED16.5) livers were purified using MACS system and hepatoblast-specific E-cadherin antibody. The cells were cultured in 100-mm dish at 75% confluence and transfected with a mixture of DNA containing 4 μg of pLentG-KOSM, 3.5 μg of pCMV-VSVG, and 2 μg of psPAX2 by Fugene HD. Twenty-four hours after transfection, the supernatant of transfected cells was collected and filtered through a 0.45 μm pore-size filter. The filtered lentiviral particles were concentrated by ultracentrifugation. For virus infection, mouse embryonic fibroblast (MEF) and mouse hepatic lineage cells were seeded in a 6-well plate at 1 × 105 cells per well one day before transduction. The medium was replaced with virus-containing supernatant, incubated for 2-3 hr and then cultured up to 7days with fresh media. For iPSC induction, the infected cells were trypsinized at day 7, plated in 1:5 ratio into six well plates containing feeder cells and incubated until appearance of ES-like cells.
|
| Growth protocol |
All experiments were conducted according to the National Institutes of Health (NIH) guidelines for animal care and following a study protocol approved by the National Cancer Institute (NCI) Animal Care and Use Committee. Mice had free access to food and water.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Quiagen Rneasy Kit. Quality control and measurement were performed using NanoDrop and Agilent Bioanalyser.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared with the Ambion Totalprep RNA Amplification kit for Illumina arrays
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
Donor Cells
replicate 1
|
| Data processing |
Data was processed by using Gene Expression Module v 1.1.1 in Genome Studio V2009.1 from illumina, Inc. The background noise was substrated and quantile normalization was applied.
|
| |
|
| Submission date |
Oct 20, 2011 |
| Last update date |
Oct 02, 2012 |
| Contact name |
Daekwan Seo |
| E-mail(s) |
daekwan_seo@nih.gov
|
| Phone |
301-496-5688
|
| Fax |
301-496-0734
|
| Organization name |
NIH
|
| Department |
NCI
|
| Lab |
LEC
|
| Street address |
37 Convent Dr. Rm 4140
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20878 |
| Country |
USA |
| |
|
| Platform ID |
GPL6885 |
| Series (1) |
| GSE33110 |
Differentiation stage-specific donor memory in induced pluripotent stem cells (iPSC) generated from hepatic lineage cells |
|
