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


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 - [Gdanski Uniwersytet Medyczny]
Alina Mieczkowska,,
- Gdanski Uniwersytet Medyczny
, Adriana Schumacher - [Gdanski Uniwersytet Medyczny]
Adriana Schumacher,,
- Gdanski Uniwersytet Medyczny
, Natalia Filipowicz - [Gdanski Uniwersytet Medyczny]
Natalia Filipowicz,,
- Gdanski Uniwersytet Medyczny
, Anna Wardowska - [Gdanski Uniwersytet Medyczny]
Anna Wardowska,,
- Gdanski Uniwersytet Medyczny
, Maciej Zieliński - [Gdanski Uniwersytet Medyczny]
Maciej Zieliński,,
- Gdanski Uniwersytet Medyczny
, Piotr Madanecki - [Gdanski Uniwersytet Medyczny]
Piotr Madanecki,,
- Gdanski Uniwersytet Medyczny
, Ewa Nowicka - [Gdanski Uniwersytet Medyczny]
Ewa Nowicka,,
- Gdanski Uniwersytet Medyczny
, Paulina Langa - [Gdanski Uniwersytet Medyczny]
Paulina Langa,,
- Gdanski Uniwersytet Medyczny
, Milena Deptuła - [Gdanski Uniwersytet Medyczny]
Milena Deptuła,,
- Gdanski Uniwersytet Medyczny
, Jacek Zieliński - [Gdanski Uniwersytet Medyczny]
Jacek Zieliński,,
- Gdanski Uniwersytet Medyczny
et al.`
Journal seriesScientific Reports, ISSN 2045-2322, (A 40 pkt)
Issue year2018
Publication size in sheets0.6
Article number11339
ASJC Classification1000 Multidisciplinary
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)40
Score sourcejournalList
ScoreMinisterial score = 40.0, 29-05-2020, ArticleFromJournal
Publication indicators WoS Citations = 1; Scopus Citations = 7; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 1.401; WoS Impact Factor: 2018 = 4.011 (2) - 2018=4.525 (5)
Citation count*12 (2020-05-30)
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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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