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Sample GSM5813590 Query DataSets for GSM5813590
Status Public on Jan 13, 2022
Title Lipopolysaccharides 24h induced DPSCs + Laser irradiated DPSCs replicate 1
Sample type RNA
 
Source name Human Dental Pulp Stem Cells
Organism Homo sapiens
Characteristics cell type: Stem Cells
protocol: Lipopolysaccharides 24h induced + Laser irradiated
tissue: pulp
Treatment protocol To promote in vitro cellular inflammatory response, 2 ×106 of DPSCs were seeded in six wells plates with α-MEM, supplemented with 10% FCS, 100 μML-ascorbic acid, 2mML-glutamine and penicillin (100U/mL)/streptomycin (100 mg/ml) and incubated from 24 h at 37°C with 5% CO2 for complete adhesion. Previous to the experiment cells were starved for 6hs in αMEM without FCS. After starvation period, 10 μg/ml of LPS were added in α-MEM, supplemented with 10% FCS, 100 μML-ascorbic acid, 2 mML-glutamine and penicillin (100 U/mL)/streptomycin (100 mg/ml), for 24 hs.Part of these cells was further irradiated with a laser device in the infrared wavelength (808 nm) that delivered a total energy of 6 J (100mW× 60 s). (Group 1); while the other part remained not irradiated for subsequent analysis (Group 2).
Growth protocol The cells were seeded into culture flasks with α-MEM, supplemented with 10% FCS, 100 μML-ascorbic acid, 2mML-glutamine and penicillin (100 U/mL)/streptomycin (100 mg/ml) and incubated at 37°C with 5% CO2. In the initial stage, after adhering to culture flasks, cells grew slowly. Cell colonies were identified after 10–14 days, with their fibroblastoid appearance, which showed to be dependent on the cell density obtained in initial plating. After reach 70% of confluence, cells were considered ready to proceed with experiments.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted with RNeasy Mini Kit followed by DNase I treatment.
Label SYBR Green
Label protocol The Qiagen RT² Profiler PCR Arrays are highly reliable and sensitive gene expression profiling tools for analyzing focused panels of genes in signal transduction, biological processes or disease research pathways using real-time PCR. Each cataloged RT² Profiler PCR Array contains a list of the pathway-focused genes as well as five housekeeping (reference) genes on the array. In addition, each array contains a panel of proprietary controls to monitor genomic DNA contamination (GDC) as well as the first strand synthesis (RTC) and real-time PCR efficiency (PPC). The qPCR Assays used in PCR Arrays are laboratory-verified and optimized to work under standard conditions enabling a large number of genes to be assayed simultaneously. Their specificity is guaranteed when RT² SYBR Green qPCR Master Mixes are used as part of the complete PCR Array System protocol.
 
Hybridization protocol n/a
Scan protocol n/a
Description Group 2
SAMPLE 5
Data processing CT values were exported to an Excel file to create a table of CT values. This table was then uploaded on to the data analysis web portal at http://www.qiagen.com/geneglobe
Relative quantification was carried out using the ΔCt method. This method results in ratios between the target genes and the housekeeping reference gene, in this case, the enzyme β-2-microglobulin (positive control) for DSPCs. This assay is based on gene fold-Regulation approach, that represents in fold-change (fc) results of the evaluated genes. Fold-change values greater than one indicates a positive- or an upregulation in gene expression, while fold-change values less than one indicate a negative or down-regulation, of the evaluated gene.
The p-values were calculated based on a Student’s t-test of the replicate 2^(ΔCt) values for each gene in the experimental or control groups
 
Submission date Jan 12, 2022
Last update date Jan 13, 2022
Contact name Fabio Dupart Nascimento
E-mail(s) fdnascimento@gmail.com
Phone 11985584947
Organization name Federal University of Sao Paulo
Department Biochemistry
Street address Rua Doutor Gabriel Piza, 636 apto 23
City São Paulo
State/province SP
ZIP/Postal code 02036-011
Country Brazil
 
Platform ID GPL31218
Series (1)
GSE193448 Real-time quantitative PCR analysis of human dental pulp stem cells

Supplementary data files not provided
Processed data are available on Series record

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