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

Alina Mieczkowska , Adriana Schumacher , Natalia Filipowicz , Anna Wardowska , Maciej Zieliński , Piotr Madanecki , Ewa Nowicka , Paulina Langa , Milena Deptuła , Jacek Zieliński , Karolina Kondej , Alicja Renkielska , Patrick G. Buckley , David K. Crossman , Michael R. Crowley , Artur Czupryn , Piotr Mucha , Paweł Sachadyn , Łukasz Janus , Piotr Skowron , Sylwia Rodziewicz-Motowidło , Mirosława Cichorek , Michał Pikuła , Arkadiusz Piotrowski

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.
Author Alina Mieczkowska
Alina Mieczkowska,,
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, Adriana Schumacher
Adriana Schumacher,,
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, Natalia Filipowicz
Natalia Filipowicz,,
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, Anna Wardowska
Anna Wardowska,,
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, Maciej Zieliński
Maciej Zieliński,,
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, Piotr Madanecki
Piotr Madanecki,,
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, Ewa Nowicka
Ewa Nowicka,,
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, Paulina Langa
Paulina Langa,,
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, Milena Deptuła
Milena Deptuła,,
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, Jacek Zieliński
Jacek Zieliński,,
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et al.`
Journal seriesScientific Reports, ISSN 2045-2322, (A 40 pkt)
Issue year2018
Vol8
Pages1-13
Publication size in sheets0.6
URL https://www.nature.com/articles/s41598-018-29477-5.pdf
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)40
ScoreMinisterial score = 40.0, 21-08-2018, ArticleFromJournal
Ministerial score (2013-2016) = 40.0, 21-08-2018, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 4.259 (2) - 2016=4.847 (5)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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