|
| Status |
Public on Oct 04, 2008 |
| Title |
Human dermal lymphatic endothelial cell 2 |
| Sample type |
RNA |
| |
|
| Source name |
Human primary dermal microvascular lymphatic endothelial cell (LEC) isolated from neonatal human foreskin
|
| Organism |
Homo sapiens |
| Characteristics |
CD45-/CD34-/CD31+ Human Lymphatic Endothelial Cell (LEC)
|
| Growth protocol |
Cells were propagated in endothelial cell basal medium (Cambrex Corp.; East Rutherford, NJ ) containing 20% fetal bovine serum, antibiotics, 2 mmol/L L-glutamine (Invitrogen Corp.; Carlsbad, CA), 10 µg/mL hydrocortisone acetate and 2.5 10-2 mg/ml N-6,2’-O-dibutyryl adenosine-3,5’-cyclic monophosphate (Sigma-Aldrich; St. Louis, MO).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Isolate total cellular RNA with 1 ml of the Trizol reagent (Invitrogen) after washing one 80-90 % confluent 10 cm culture dish with PBS. Homogenize the sample using a cell scraper and add 0.2 ml of chloroform in a 1.5 ml tube. Shake the sample vigorously for 15 seconds and allow standing for 5 minutes. Centrifuge the resulting mixture at 12,000 x g for 15 minutes at 4 degree Celsius. Transfer the aqueous phase to a fresh tube and add 0.5 ml of iso-propanol and mix. Allow the sample to stand for 5 minutes at room temperature and centrifuge at 12,000 x g for 10 minutes at 4 degrees Celsius. Remove the supernatant and wash the RNA pellet by adding 1 ml of 75% ethanol. Vortex the sample and then centrifuge at 12,000 x g for 5 minutes at 4 degrees Celsius. Remove the ethanol and briefly dry the RNA pellet for 5 minutes and add 50 micro-liter of RNAase/DNAase free water and mix by repeated pipetting until RNA pellet is fully dissolved.
|
| Label |
TaqMan Probe
|
| Label protocol |
TaqMan™Quantitative RT-PCR - FAM
|
| |
|
| Hybridization protocol |
TaqMan™Quantitative RT-PCR
|
| Scan protocol |
TaqMan™ RT-PCR - Fluorescence
|
| Description |
LEC2
|
| Data processing |
Raw data were normalized to the cycle number (Ct) of beta-Actin expression
|
| |
|
| Submission date |
Apr 30, 2008 |
| Last update date |
Jul 25, 2008 |
| Contact name |
Jay Woo Shin |
| E-mail(s) |
jay.shin@gsc.riken.jp
|
| Organization name |
ETH Zurich
|
| Department |
Institute of Pharmaceutical Sciences
|
| Lab |
Michael Detmar
|
| Street address |
Wolfgang-Paulistrasse 10; HCI H394
|
| City |
Zurich |
| State/province |
Zurich |
| ZIP/Postal code |
8093 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL6802 |
| Series (2) |
| GSE11306 |
Quantification of vascular lineage-specific differentiation (cell type comparison) |
| GSE11308 |
Quantification of vascular lineage-specific differentiation |
|
