Assessment of DNA Damage in Human Bone Marrow Cells and Multipotent Mesenchymal Stromal Cells

Nikitina, V. A.; Chausheva, A. I.; Жанатаев, А. К.; Osipova, E. Yu.; Дурнев, А. Д.; Бочков, Н. П.

doi:10.1007/s10517-011-1379-8

Cited by 6 publications

(5 citation statements)

References 9 publications

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“…Froelich et al (2013) also detected chromosomal aberrations by means of the chromosomal aberration test, even when the comet assay did not detect damage. This same finding is also reported by Nikitina et al (2011) andBochkov et al (2006).…”

Section: Discussionsupporting

confidence: 90%

Evaluation of pH effects on genomic integrity in adipose-derived mesenchymal stem cells using the comet assay

et al. 2015

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ABSTRACT. The use of mesenchymal stem cells (MSCs) in experimental, clinical, and therapeutic trials has grown in recent years. However, the issue remains of whether these procedures are completely safe for transplant patients. Therefore, this study was designed and carried out with the aim of evaluating two different comet assay protocols for genomic damage pattern analysis in MSCs derived from adipose tissue. The analyzed and interpreted results suggest that genetic testing is needed to support clonal expansion safety in cell therapy procedures with MSCs. Furthermore, they also suggest that if the comet assay technique would be used as a genomic integrity screening assay, the protocol performed at pH = 12 (that yielded a frequency of damaged cells: tail intensity = 9.50 ± 0.60, tail moment = 0.0122 ± 0.0007; results are reported as means ± standard deviation) would be indicated as genomic damage, and that subsequent single-strand breaks occur at pH > 13 (frequency of damaged cells: tail intensity = 30.71 ± 4.23, tail moment = 0.0447 ± 0.0073). Our study demonstrates that, in the era of regenerative medicine, it is necessary to standardize and establish a battery of tests in order to identify genomic damage prior to MSC transplantation.

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Section: Discussionsupporting

confidence: 90%

Evaluation of pH effects on genomic integrity in adipose-derived mesenchymal stem cells using the comet assay

et al. 2015

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“…However, studies for hMSCs report low levels of chromosomal aberrations among donor samples [15, 16] and during culture expansion [5, 17, 18]. Low levels of non-clonal chromosomal aberrations in MSCs used in clinical trials have been observed, but there are no reports of malignant transformation [19].…”

Section: Introductionmentioning

confidence: 99%

Chromosomal stability of mesenchymal stromal cells during in vitro culture

et al. 2016

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Background aims Mesenchymal stromal cells (MSCs) are being investigated for use in cell therapy. The extensive in vitro expansion necessary to obtain sufficient cells for clinical use increases the risk that genetically abnormal cells will arise and be propagated during cell culture. Genetic abnormalities may lead to transformation and poor performance in clinical use, and are a critical safety concern for cell therapies using MSCs. Methods We used spectral karyotyping (SKY) to investigate the genetic stability of human MSCs from ten donors during passaging. Results Our data indicate that chromosomal abnormalities exist in MSCs at early passages and can be clonally propagated. The karyotypic abnormalities observed during our study diminished during passage. Conclusions Karyotyping of MSCs reveals characteristics which may be valuable in deciding the suitability of cells for further use. Karyotypic analysis is useful for monitoring the genetic stability of MSCs during expansion.

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“…Indeed, they must be produced via a robust GMP process to ensure safety and efficacy: the validation of the process is a critical step, considering that living cells are bioresponsive and can rapidly react to changes of the culture environment. In particular, both MSC and ASC in culture are subject to genomic and/or chromosomal mutations and the karyotype can change (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25). It has not yet been clarified in what culture conditions the mutations are most frequent, and the results obtained by independent research groups are often contradictory, as reviewed by Ferreira in 2012 (26).…”

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confidence: 99%

Analysis of the Karyotype of Expanded Human Adipose-Derived Stem Cells for Bone Reconstruction of the Maxillo-Facial Region

Bellotti

Stanco

Ragazzini

et al. 2013

Int J Immunopathol Pharmacol

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Mesenchymal stem cells (MSC) and adipose-derived stem cells (ASC) were recently proposed for bone maxillofacial reconstruction in association with biomaterials. For this application MSC must be ex-vivo expanded in order to obtain, for a given volume of implanted biomaterial, a relevant number of bone forming cells. Previously conducted pre-clinical studies suggested that a concentration of 6 x 10 8 ASC associated with 900 mg of anorganic bovine bone (ABB) could be effective for human maxillary sinus floor elevation. A keystone issue to guarantee the quality and safety of Advanced Therapy Medicinal Products containing expanded MSC and ASC is their chromosome stability in culture: this topic has been widely investigated and conflicting results have been published. Abnormal karyotype of human ex-vivo expanded MSC and ASC was found by some authors, while, at the same time, several other studies showed the MSC and ASC karyotype to be normal. It is therefore important that all the results obtained on MSC andASC karyotype analysis be published. Given this context, the aim of this manuscript, aim of this manuscript is to verify the karyotype stability of ASC in view oftheir applications in clinical trials. ASC obtained from the adipose tissue of 4 donors were expanded over extended culture time. Based on previous ASC expansions we hypothesized to be able to obtain 6 x lOS cells by passage 7. Karyotype analysis 000 metaphases was planned to be investigated at passage 2, 7, and 15 in all the cultures. No abnormalities were found in the karyotype of two donors at all the passages tested, while a translocation was found in 2 metaphases of a donor at passage 7, but not at passage 15, and in the fourth donor in 5 metaphases a trisomy was found at passage 15. Chromosomal abnormalities were detected only after extended ASC expansion. Whether these anomalies can be related to risk for the patient's safety will have to be demonstrated by in-vivo studies.

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Assessment of DNA Damage in Human Bone Marrow Cells and Multipotent Mesenchymal Stromal Cells

Cited by 6 publications

References 9 publications

Evaluation of pH effects on genomic integrity in adipose-derived mesenchymal stem cells using the comet assay

Evaluation of pH effects on genomic integrity in adipose-derived mesenchymal stem cells using the comet assay

Chromosomal stability of mesenchymal stromal cells during in vitro culture

Analysis of the Karyotype of Expanded Human Adipose-Derived Stem Cells for Bone Reconstruction of the Maxillo-Facial Region

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