|
| Status |
Public on Jun 29, 2022 |
| Title |
HA-convective flow control 33 |
| Sample type |
RNA |
| |
|
| Source name |
Human astrocytes from healthy normal brain; Cell Applications, Inc.
|
| Organism |
Homo sapiens |
| Characteristics |
treatment group: conP
dcs current (ma): n/a
rna collection time: immediate
|
| Treatment protocol |
Each experiment consisted of four groups: conP, DCS P, conS, and DCS S. Direct Current Stimulation was applied with (DCS P) or without (DCS S) a pressure differential that induced convective fluid flow across the cell monolayer. Monolayers exposed only to convective flow (conP) were also examined. A static control (conS) kept in an incubator for the duration of the experiment was used as the calibrator sample for gene expression analysis.
|
| Growth protocol |
Human Astrocytes were grown according to the manufacturer's instructions and plated at 3x104 cells/cm2 in Transwell filters coated with fibronectin and cultured for 5-6 days before DCS experiments.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was collected either immediately after DCS, or 1 hour after DCS using the RNeasy Mini kit from Qiagen.
|
| Label |
SYBR Green
|
| Label protocol |
Total RNA was reversed transcribed using the High-capacity cDNA Reverse Transcription kit from Thermo Scientific. Real time RT-qPCR was performed on the 7300 Real Time PCR System from Applied Biosystems with 45 cycles at 95 oC for 20 seconds, 60 oC for 30 seconds, and 72 oC for 30 seconds.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
HA-convective flow control
test
|
| Data processing |
Raw Ct values were obtained from the Applied Biosystems 7300 Real Time PCR System as .csv files and data was processed in Excel.
Matrix non-normalized reports the average Ct value of duplicate samples.
Target signal normalized against the average of two housekeeping genes: GAPDH and ACTB.
Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target − Ct_HKG)]
Fold Change worksheet reports test/control ratios. For each experiment, test samples were normalized to the corresponding control (conS) sample.
|
| |
|
| Submission date |
Jun 28, 2022 |
| Last update date |
Jun 29, 2022 |
| Contact name |
Limary Cancel |
| Organization name |
The City College of New York
|
| Department |
Biomedical Engineering
|
| Street address |
160 Convent Ave
|
| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10031 |
| Country |
USA |
| |
|
| Platform ID |
GPL32410 |
| Series (1) |
| GSE207140 |
Real-time quantitative PCR analysis of human astrocytes exposed to pressure-driven flow and/or Direct Current Stimulation in vitro |
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