|
| Status |
Public on May 12, 2010 |
| Title |
HMEC 184D14pM85X.1 Stasis |
| Sample type |
RNA |
| |
|
| Source name |
HMEC
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: mammary epithelial cell
individual: 184
passage: 14p
growth status: Stasis
|
| Treatment protocol |
Cell cultures grown in the indicated media were harvested and different points in their growth from early passage good growing to near stasis, agonescence, or senescence.
|
| Growth protocol |
Finite life span HMEC and HMFC from Specimens 184, 48 and 240L were obtained from reduction mammoplasty tissue of women aged 21, 16, and 19 respectively. HMEC were grown starting from organoids in either serum free MCDB170 medium, or the serum-containing M85+CT±X medium described in this publication. HMFC were grown in DME/F12+FCS+I
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cell were lysed with trizol reagent (Invitrogen) according manufacturer's protocol. RNA was resuspended in 100 µl nuclease free water and subject to a second step purification on ion exchange column (Qiagen)
|
| Label |
Biotin
|
| Label protocol |
For each sample, the RNA target is prepared by annealing 2.0µg of total RNA in 5µl water with 5µl of 10µM T7 (Oligo dt) primer. First and second strand cDNA synthesis is performed and the cDNA products purified using magnetic beads. Labeled cRNA is synthesized from the cDNA using T7 RNA polymerase followed by clean-up with magnetic beads. Purified cRNA is quantitated and fragmented.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip HT_HG-U133A Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Human Cell Lines
|
| Data processing |
Raw probe level intensities were processed for background correction, normalization and summarization using Probel level linear (PLM) model in LIMMA module(v.2.16.3) of Bioconductor package.
|
| |
|
| Submission date |
May 12, 2009 |
| Last update date |
May 12, 2009 |
| Contact name |
Sanchita Bhattacharya |
| E-mail(s) |
SBhattacharya@lbl.gov
|
| Phone |
510-486-6881
|
| Organization name |
MIT
|
| Street address |
1 Cyclotron Rd
|
| City |
Berkely |
| State/province |
CA |
| ZIP/Postal code |
94720 |
| Country |
USA |
| |
|
| Platform ID |
GPL3921 |
| Series (1) |
| GSE16058 |
Distinctions between the stasis and telomere attrition senescence barriers in cultured human mammary epithelial cells |
|
