Immunophenotyping and transcriptional profiling of in vitro cultured human adipose tissue derived stem cells.

Autor: Mieczkowska A; Faculty of Pharmacy, Medical University of Gdansk, Gdansk, Poland., Schumacher A; Department of Embryology, Faculty of Medicine, Medical University of Gdansk, Gdansk, Poland., Filipowicz N; Faculty of Pharmacy, Medical University of Gdansk, Gdansk, Poland., Wardowska A; Department of Clinical Immunology and Transplantology, Medical University of Gdansk, Gdansk, Poland.; Laboratory of Tissue Engineering and Regenerative Medicine, Department of Embryology, Faculty of Medicine, Medical University of Gdansk, Gdansk, Poland., Zieliński M; Department of Clinical Immunology and Transplantology, Medical University of Gdansk, Gdansk, Poland., Madanecki P; Faculty of Pharmacy, Medical University of Gdansk, Gdansk, Poland., Nowicka E; Department of Clinical Anatomy, Medical University of Gdansk, Gdansk, Poland., Langa P; Department of Clinical Immunology and Transplantology, Medical University of Gdansk, Gdansk, Poland., Deptuła M; Department of Embryology, Faculty of Medicine, Medical University of Gdansk, Gdansk, Poland.; Laboratory of Tissue Engineering and Regenerative Medicine, Department of Embryology, Faculty of Medicine, Medical University of Gdansk, Gdansk, Poland., Zieliński J; Department of Surgical Oncology, Medical University of Gdansk, Gdansk, Poland., Kondej K; Department of Plastic Surgery, Medical University of Gdansk, Gdansk, Poland., Renkielska A; Department of Plastic Surgery, Medical University of Gdansk, Gdansk, Poland., Buckley PG; GMI Genomics Centre, Genomics Medicine Ireland, Dublin, Ireland., Crossman DK; Heflin Center for Genomic Sciences, University of Alabama at Birmingham, Birmingham, Alabama, USA., Crowley MR; Heflin Center for Genomic Sciences, University of Alabama at Birmingham, Birmingham, Alabama, USA., Czupryn A; Laboratory of Neurobiology, Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology PAS, Warsaw, Poland., Mucha P; Department of Biochemistry, Faculty of Chemistry, University of Gdansk, Gdansk, Poland., Sachadyn P; Laboratory for Regenerative Biotechnology, Gdansk University of Technology, Gdansk, Poland., Janus Ł; MedVentures company sp. z o.o., Poznań, Poland., Skowron P; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk, Poland., Rodziewicz-Motowidło S; Department of Biomedicinal Chemistry, Faculty of Chemistry, University of Gdansk, Gdansk, Poland., Cichorek M; Department of Embryology, Faculty of Medicine, Medical University of Gdansk, Gdansk, Poland., Pikuła M; Department of Clinical Immunology and Transplantology, Medical University of Gdansk, Gdansk, Poland. pikula@gumed.edu.pl.; Laboratory of Tissue Engineering and Regenerative Medicine, Department of Embryology, Faculty of Medicine, Medical University of Gdansk, Gdansk, Poland. pikula@gumed.edu.pl., Piotrowski A; Faculty of Pharmacy, Medical University of Gdansk, Gdansk, Poland. arpiotr@gumed.edu.pl.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2018 Jul 27; Vol. 8 (1), pp. 11339. Date of Electronic Publication: 2018 Jul 27.
DOI: 10.1038/s41598-018-29477-5
Abstrakt: 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.
Databáze: MEDLINE
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