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Immunophenotyping and transcriptional profiling of in vitro cultured human adipose tissue derived stem cells
Abstract
Adipose-derived stem cells (ASCs) have become an important research model in regenerative medicine. However, there are controversies regarding the impact of prolonged cell culture on the ASCs phenotype and their differentiation potential. Hence, we studied 10 clinical ASCs replicates from plastic and oncological surgery patients, in six-passage FBS supplemented cultures. We quantified basic mesenchymal cell surface marker transcripts and the encoded proteins after each passage. In parallel, we investigated the differentiation potential of ASCs into chondrocytes, osteocytes and adipocytes. We further determined the effects of FBS supplementation and subsequent deprivation on the whole transcriptome by comprehensive mRNA and miRNA sequencing. Our results show that ASCs maintain differentiation potential and consistent profile of key mesenchymal markers, with apparent expression of distinct isoforms, in long-term cultures. No significant differences were observed between plastic and oncological surgery cohorts. ASCs in FBS supplemented primary cultures are almost committed to mesenchymal lineages as they express key epithelial-mesenchymal transition genes including early mesenchymal markers. Furthermore, combined mRNA/miRNA expression profiling strongly supports a modulatory role for the miR-30 family in the commitment process to mesenchymal lineages. Finally, we propose improvements to existing qPCR based assays that address alternative isoform expression of mesenchymal markers.
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Authors (24)
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Alina Mieczkowska
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Adriana Schumacher mgr
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Natalia Filipowicz
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Anna Wardowska
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Maciej Zieliński
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Piotr Madanecki
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Ewa Nowicka
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Paulina Langa
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Milena Deptuła
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Jacek Zieliński dr hab.
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Karolina Kondej
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Alicja Renkielska
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Patrick Buckley
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David Crossman
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Michael Crowley
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Artur Czupryn dr
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Piotr Mucha dr hab.
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Łukasz Janus dr
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Piotr Skowron prof. dr hab.
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Sylwia Rodziewicz-Motowidło dr hab.
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Mirosława Cichorek
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Michał Pikuła dr hab. med.
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Arkadiusz Piotrowski dr hab.
Cite as
Full text
- Publication version
- Accepted or Published Version
- DOI:
- Digital Object Identifier (open in new tab) 10.1038/s41598-018-29477-5
- License
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Keywords
Details
- Category:
- Articles
- Type:
- artykuł w czasopiśmie wyróżnionym w JCR
- Published in:
-
Scientific Reports
no. 8,
edition 1,
pages 1 - 13,
ISSN: 2045-2322 - Language:
- English
- Publication year:
- 2018
- Bibliographic description:
- Mieczkowska A., Schumacher A., Filipowicz N., Wardowska A., Zieliński M., Madanecki P., Nowicka E., Langa P., Deptuła M., Zieliński J., Kondej K., Renkielska A., Buckley P., Crossman D., Crowley M., Czupryn A., Mucha P., Sachadyn P., Janus Ł., Skowron P., Rodziewicz-Motowidło S., Cichorek M., Pikuła M., Piotrowski A.: Immunophenotyping and transcriptional profiling of in vitro cultured human adipose tissue derived stem cells// Scientific Reports. -Vol. 8, (2018), s.1-13
- DOI:
- Digital Object Identifier (open in new tab) 10.1038/s41598-018-29477-5
- Sources of funding:
- Verified by:
- Gdańsk University of Technology
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