Chromosomal aberrations and deoxyribonucleic acid single-strand breaks in adipose-derived stem cells during long-term expansion in vitro |
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| Authors: | Katrin Froelich Johannes Mickler Gudrun Steusloff Antje Technau Mario Ramos Tirado Agmal Scherzed Stephan Hackenberg Andreas Radeloff Rudolf Hagen Norbert Kleinsasser |
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| Affiliation: | 1. Pharmaceutical Production Research Facility, Schulich School of Engineering, University of Calgary, Calgary, Alberta, Canada;2. McCaig Institute for Bone and Joint Health, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada;3. Science for Life Laboratory, Royal Institute of Technology, SE-17121 Solna, Sweden;4. School of Biotechnology, AlbaNova University Center, Royal Institute of Technology, SE-106 91 Stockholm, Sweden;6. Department of Medical Epidemiology, Karolinska Institute, SE-171 77 Stockholm, Sweden;12. Division of Pharmacoproteomics, National Cancer Center Research Institute, Tokyo 104-0045, Japan;1. Department of Urology, University of Antwerp, Antwerp, Belgium;2. Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Antwerp, Belgium;3. StatUA Centre for Statistics, University of Antwerp, Antwerp, Belgium;4. Laboratory of Experimental Surgery, Antwerp Surgical Training and Research Center, University of Antwerp, Antwerp, Belgium;5. Department of Pathology, University Hospital Antwerp, Antwerp, Belgium |
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| Abstract: | Background aimsAdipose-derived stem cells (ASCs) are a promising mesenchymal cell source for tissue engineering approaches. To obtain an adequate cell amount, in vitro expansion of the cells may be required in some cases. To monitor potential contraindications for therapeutic applications in humans, DNA strand breaks and chromosomal aberrations in ASCs during in vitro expansion were examined.MethodsAfter isolation of ASC from human lipoaspirates of seven patients, in vitro expansion over 10 passages was performed. Cells from passages 1, 2, 3, 5 and 10 were used for the alkaline single-cell microgel electrophoresis (comet) assay to detect DNA single-strand breaks and alkali labile as well as incomplete excision repair sites. Chromosomal changes were examined by means of the chromosomal aberration test.ResultsDuring in vitro expansion, ASC showed no DNA single-strand breaks in the comet assay. With the chromosomal aberration test, however, a significant increase in chromosomal aberrations were detected.ConclusionsThe study showed that although no DNA fragmentation could be determined, the safety of ASC cannot be ensured with respect to chromosome stability during in vitro expansion. Thus, reliable analyses for detecting ASC populations, which accumulate chromosomal aberrations or even undergo malignant transformation during extensive in vitro expansion, must be implemented as part of the safety evaluation of these cells for stem cell–based therapy. |
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