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Status |
Public on Mar 13, 2018 |
Title |
MSC1_in hPL-gel_with_TN [expression] |
Sample type |
RNA |
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Source name |
MSCs, within hPL-gel with TN
|
Organism |
Homo sapiens |
Characteristics |
donor: 1 cell type: mesenchymal stell cells (MSCs) passage: 5 culture condition: within hPL-gel with thrombin (TN)
|
Treatment protocol |
To determine the impact of culture conditions on gene expression of primary MSCs, we treated the cells for three weeks in parallel with the following conditions: i) MSC were cultured on tissue culture plastic (TCP) in standard medium supplemented with 10% human platelet lysate (hPL-medium); ii) on TCP in hPL-plasma (after depletion of coagulation factors); iii) on hPL-gel generated without thrombin (TN), iv) on hPL-gel generated with TN (Sigma Aldrich, Hamburg, Germany), or iv) by trapping of the cells within hPL-gel with addition of TN. Alternatively, iPSCs were differentiated toward MSCs with the following conditions: iPSCs were either seeded on TCP with i) MSC standard medium supplemented with 10% human platelet lysate (hPL-medium) or ii) hPL-plasma (after depletion of coagulation factors); or on hPL-gel generated iii) without thrombin (TN) or iv) with TN (Sigma Aldrich, Hamburg, Germany) for a period of 35 days.
|
Growth protocol |
MSCs were isolated from bone marrow after written consent using guidelines approved by the Ethic Committee on the Use of Human Subjects at the University of Aachen (permit number: EK128/09). For some of the profiles, the MSCs were reprogrammed into iPSCs. To this end, MSCs of three different donors (passage 2) were infected with pCXLE-based episomal plasmids (Addgene, Cambridge, MA, USA) carrying the OCT3/4, SOX2, KLF4, LIN28 and c-MYC genes. Established iPSCs were cultured on TCP coated with vitronectin (0.5 µg/cm²) in StemMACS iPS-Brew XF (all Miltenyi Biotec GmbH, Bergisch Gladbach, Germany).
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was isolated using the Macherey-Nagel RNA Plus kit.
|
Label |
biotin
|
Label protocol |
Standard Illumina protocol.
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|
|
Hybridization protocol |
Standard Illumina protocol.
|
Scan protocol |
Standard Illumina protocol.
|
Data processing |
Raw data were quantile normalized using the GenomeStudio Software (Illumina, San Diego, USA). For comparison of gene expression of iMSCs and MSCs, all transcripts with a detection p > 0.01 were excluded.
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Submission date |
Feb 18, 2017 |
Last update date |
Mar 13, 2018 |
Contact name |
Wolfgang Wagner |
E-mail(s) |
wwagner@ukaachen.de
|
Phone |
+49 241 8088611
|
Organization name |
RWTH Aachen University
|
Department |
Helmholtz Institute for Biomedical Engineering
|
Lab |
Stem Cell Biology and Cellular Engineering
|
Street address |
Pauwelsstrasse 20
|
City |
Aachen |
ZIP/Postal code |
52074 |
Country |
Germany |
|
|
Platform ID |
GPL10558 |
Series (2) |
GSE95060 |
Soft Hydrogels Support Differentiation of Induced Pluripotent Stem Cells toward Mesenchymal Stromal Cells [expression] |
GSE95061 |
Soft Hydrogels Support Differentiation of Induced Pluripotent Stem Cells toward Mesenchymal Stromal Cells |
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