|
| Status |
Public on Oct 28, 2020 |
| Title |
E: Exs-Ad-control-3 |
| Sample type |
RNA |
| |
|
| Source name |
Ad-control transfected PDLSCs
|
| Organism |
Homo sapiens |
| Characteristics |
adenovirus: Ad-control transfected
donor: donor 3
age: 24y
tissue: periodontal ligament stem cells (PDLSCs)
|
| Treatment protocol |
Human periodontal ligament stem cells were firstly transfected with Ad-P2X7 or Ad-control. Culture these cells with a-MEM medium supplemented without fetal bovine serum for 24 hours at 37 ºC in a humidified incubator with 5% CO2.Then, we isolated the exosomes from the culture medium of these gene modified stem cells.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA quantity and quality were measured by NanoDrop ND-1000. RNA integrity was assessed by standard denaturing agarose gel electrophoresis.
|
| Label |
Cyanine 3-pCp
|
| Label protocol |
Sample labeling and array hybridization were performed according to the Agilent miRNA Microarray Systemwith miRNA Complete Labeling and Hyb Kit protocol (Agilent Technology). Briefly, total miRNA from each sample was labeled with Cyanine 3-pCp under the action of T4 RNA ligase. The labeled cRNA over the procession of inspissation and desiccation and then redissolved with water.
|
| |
|
| Hybridization protocol |
1 μg of each labeled cRNA was fragmented by adding 11 μl 10 × Blocking Agent and 2.2 μl of 25×Fragmentation Buffer, then heated at 60°C for 30 min, and finally 55 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 100 μl of hybridization solution was dispensed into the gasket slide and assembled to the gene expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven. The hybridized arrays were washed, fixed and scanned with using the Agilent Microarray Scanner (part number G2505C).
|
| Scan protocol |
Agilent Feature Extraction software (version 11.0.1.1) was used to analyze acquired array images.
|
| Description |
Gene expression of exosomes which were derived from Ad-control transfected PDLSCs.
|
| Data processing |
Quantile normalization and subsequent data processing were performed with using the GeneSpring GX v12.1 software package (Agilent Technologies). After quantile normalization of the raw data, miRNAs that at least 3 out of 6 samples have flags in Detected (“All Targets Value”) were chosen for further data analysis. Differentially expressed miRNAs with statistical significance between the two groups were identified through Volcano Plot filtering. Differentially expressed miRNAs between the two samples were identified through Fold Change filtering. Hierarchical Clustering was performed using the R scripts.
|
| |
|
| Submission date |
Nov 13, 2019 |
| Last update date |
Oct 28, 2020 |
| Contact name |
Xinyue Xu |
| E-mail(s) |
xinyuexu_1@163.com
|
| Organization name |
The Fourth Military Medical University
|
| Street address |
Changle west road
|
| City |
Xi'an |
| State/province |
Shannxi |
| ZIP/Postal code |
710032 |
| Country |
China |
| |
|
| Platform ID |
GPL25134 |
| Series (1) |
| GSE140356 |
Retrieval of inflammation-compromised periodontal ligament stem cells from dysfunction by exosomes derived from P2X7 receptor gene-modified cells |
|
