Clonal chromosomal and genomic instability during human multipotent mesenchymal stromal cells long-term culture

Autor: Stanislav Lauk-Dubitskiy, Vitaliy Brunchukov, Dariya Usupzhanova, I. Kobzeva, Victoria Nikitina, Sergey Rodin, Andrey Bushmanov, Tatiana Karaseva, Valentin Brumberg, Yulia Suchkova, Vladimir Nugis, Astrelina Ta, Aleksandr Samoilov, Ekaterina Dobrovolskaya, Anastasia Machova, Aleksandr Ostashkin, Elena Lomonosova
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Clone (cell biology)
lcsh:Medicine
Aneuploidy
Chromosomal translocation
Animal Cells
Chromosome instability
Cell Cycle and Cell Division
lcsh:Science
Cells
Cultured
Multidisciplinary
Chromosome Biology
Stem Cells
Karyotype
Hälsovetenskaper
Chromosomal Aberrations
Cell Processes
Tetrasomy
Karyotypes
Cellular Types
Research Article
Chromosome Structure and Function
Biology
Research and Analysis Methods
Genomic Instability
Chromosomes
Immunophenotyping
Polyploidy
Cytogenetics
03 medical and health sciences
Health Sciences
Genetics
medicine
Humans
Molecular Biology Techniques
Molecular Biology
Metaphase
Chromosome Aberrations
lcsh:R
Mesenchymal stem cell
Biology and Life Sciences
Chromosome
Mesenchymal Stem Cells
Cell Biology
medicine.disease
Molecular biology
030104 developmental biology
Chromosomal Translocations
Karyotyping
lcsh:Q
Departures from Diploidy
Microsatellite Repeats
Cloning
Zdroj: PLoS ONE, Vol 13, Iss 2, p e0192445 (2018)
PLoS ONE
Popis: Background aims Spontaneous mutagenesis often leads to appearance of genetic changes in cells. Although human multipotent mesenchymal stromal cells (hMSC) are considered as genetically stable, there is a risk of genomic and structural chromosome instability and, therefore, side effects of cell therapy associated with long-term effects. In this study, the karyotype, genetic variability and clone formation analyses have been carried out in the long-term culture MSC from human gingival mucosa. Methods The immunophenotype of MSC has been examined using flow cytofluorometry and short tandem repeat (STR) analysis has been carried out for authentication. The karyotype has been examined using GTG staining and mFISH, while the assessment of the aneuploidy 8 frequency has been performed using centromere specific chromosome FISH probes in interphase cells. Results The immunophenotype and STR loci combination did not change during the process of cultivation. From passage 23 the proliferative activity of cultured MSCs was significantly reduced. From passage 12 of cultivation, clones of cells with stable chromosome aberrations have been identified and the biggest of these (12%) are tetrasomy of chromosome 8. The random genetic and structural chromosomal aberrations and the spontaneous level of chromosomal aberrations in the hMSC long-term cultures were also described. Conclusions The spectrum of spontaneous chromosomal aberrations in MSC long-term cultivation has been described. Clonal chromosomal aberrations have been identified. A clone of cells with tetrasomy 8 has been detected in passage 12 and has reached the maximum size by passage 18 before and decreased along with the reduction of proliferative activity of cell line by passage 26. At later passages, the MSC line exhibited a set of cells with structural variants of the karyotype with a preponderance of normal diploid cells. The results of our study strongly suggest a need for rigorous genetic analyses of the clone formation in cultured MSCs before use in medicine.
Databáze: OpenAIRE
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