Age-dependent impairment of adipose-derived stem cells isolated from horses
Michalina Alicka , Katarzyna Kornicka-Garbowska , Katarzyna Kucharczyk , Martyna Kępska , Michael Röcken , Krzysztof Marycz
AbstractBACKGROUND: Progressive loss of cell functionality caused by an age-related impairment in cell metabolism concerns not only mature specialized cells but also its progenitors, which significantly reduces their regenerative potential. Adipose-derived stem cells (ASCs) are most commonly used in veterinary medicine as an alternative treatment option in ligaments and cartilage injuries, especially in case of high-value sport horses. Therefore, the main aim of this study was to identify the molecular alternations in ASCs derived from three age-matched horse groups: young (< 5), middle-aged (5-15), and old (> 15 years old). METHODS: ASCs were isolated from three age-matched horse groups using an enzymatic method. Molecular changes were assessed using qRT-PCR, ELISA and western blot methods, flow cytometry-based system, and confocal and scanning electron microscopy. RESULTS: Our findings showed that ASCs derived from the middle-aged and old groups exhibited a typical senescence phenotype, such as increased percentage of G1/G0-arrested cells, binucleation, enhanced β-galactosidase activity, and accumulation of γH2AX foci, as well as a reduction in cell proliferation. Moreover, aged ASCs were characterized by increased gene expression of pro-inflammatory cytokines and miRNAs (interleukin 8 (IL-8), IL-1β, tumor necrosis factor α (TNF-α), miR-203b-5p, and miR-16-5p), as well as apoptosis markers (p21, p53, caspase-3, caspase-9). In addition, our study revealed that the protein level of mitofusin 1 (MFN1) markedly decreased with increasing age. Aged ASCs also displayed a reduction in mRNA levels of genes involved in stem cell homeostasis and homing, like TET-3, TET-3 (TET family), and C-X-C chemokine receptor type 4 (CXCR4), as well as protein expression of DNA methyltransferase (DNMT1) and octamer transcription factor 3/4 (Oct 3/4). Furthermore, we observed a higher splicing ratio of XBP1 (X-box binding protein 1) mRNA, indicating elevated inositol-requiring enzyme 1 (IRE-1) activity and, consequently, increased endoplasmic reticulum (ER) stress. We also observed reduced levels of glucose transporter 4 (GLUT-4) and insulin receptor (INSR) which indicated impaired insulin sensitivity. CONCLUSIONS: Obtained data suggest that ASCs derived from horses older than 5 years old exhibited several molecular alternations which markedly limit their regenerative capacity. The results provide valuable information that allows for a better understanding of the molecular events occurring in ASCs in the course of aging and may help to identify new potential drug targets to restore their regenerative potential.
|Journal series||Stem Cell Research & Therapy, [Stem Cell Research and Therapy], ISSN 1757-6512, (N/A 100 pkt)|
|Publication size in sheets||0.95|
|Keywords in English||Aging, Endoplasmic reticulum stress, Equine adipose-derived mesenchymal stem cells, Insulin resistance, Pro-inflammatory cytokines|
|ASJC Classification||; ; ;|
|License||Journal (articles only); published final; ; with publication|
|Score||= 100.0, 14-09-2020, ArticleFromJournal|
|Publication indicators||= 1; = 2; : 2017 = 1.266; : 2018 = 4.627 (2) - 2018=5.363 (5)|
|Citation count*||2 (2020-09-21)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.