Methodological Guidelines for Genetic Safety Testing of Cell Transplants

Бочков, Н. П.; Nikitina, V. A.; Voronina, E. S.; Кулешов, Н. П.

doi:10.1007/s10517-010-0793-7

Cited by 9 publications

(11 citation statements)

References 16 publications

(19 reference statements)

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“…Hybridization mixture for two-color FISH contained samples for three chromosome pairs: X and Y, 1 and 8, 6 and 11. Analysis was carried out as described previously [3], a total of 500 interphase nuclei were analyzed for each chromosome pair three times. Cells with one or three signals by one chromosome and two signals by the other (for pairs 1 and 8, 6 and 11), without signals or with two signals by one chromosome and one signal by the other chromosome (for X and Y pair) were referred to aneuploid.…”

Section: Methodsmentioning

confidence: 99%

“…The use of nanoparticles (NP) is a promising technology for the manufacturing cosmetic articles, drugs, biomimetics, lubricants, and fuel additives, protective and fortifying fi lms, composite, textile, packing materials, lacquers and paints [3]. The creation of nanoindustry is a priority trend of economy in Russia; carbon NP is one of the priorities in the development of materials [1,6].…”

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confidence: 99%

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Genotoxicity of Single-Walled Carbon Nanotubes: In Vitro Study on Human Embryonic Fibroblast Cells

et al. 2015

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The effects of single-walled carbon nanotubes on the levels of DNA aberrations, chromosome and genome disorders were studied on human embryonic fibroblasts, their karyotype was analyzed by the spectral karyotyping method. The level of DNA aberrations increased after 3-h exposure to the nanotubes. No appreciable increase in the incidence of aberrant metaphases, micronuclei, and chromosome 1, 6, 8, 11, X, and Y aneuploidy after 24- and 48-h incubation with the nanotubes were detected.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Genotoxicity of Single-Walled Carbon Nanotubes: In Vitro Study on Human Embryonic Fibroblast Cells

et al. 2015

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“…ND, not detected; ns, not significant stable during culturing in vitro [42][43][44][45][46][47][48]. On the contrary, abundant recent studies have revealed that human MSCs exhibited spontaneous genomic alternation with increased passage number [49][50][51][52][53][54]. Previous chromosomal profiling studies also provide that karyotypical abnormalities and chromosomal instability in early passaged human MSCs disappear during the extended passage [51,[54][55][56].…”

Section: Discussionmentioning

confidence: 99%

A model study for the manufacture and validation of clinical-grade deciduous dental pulp stem cells for chronic liver fibrosis treatment

et al. 2020

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Background: Human deciduous pulp stem cells (hDPSCs) have remarkable stem cell potency associated with cell proliferation, mesenchymal multipotency, and immunosuppressive function and have shown beneficial effects in a variety of animal disease models. Recent studies demonstrated that hDPSCs exhibited in vivo anti-fibrotic and antiinflammatory action and in vivo hepatogenic-associated liver regeneration, suggesting that hDPSCs may offer a promising source with great clinical demand for treating liver diseases. However, how to manufacture ex vivo largescale clinical-grade hDPSCs with the appropriate quality, safety, and preclinical efficacy assurances remains unclear. Methods: We isolated hDPSCs from human deciduous dental pulp tissues formed by the colony-forming unitfibroblast (CFU-F) method and expanded them under a xenogeneic-free and serum-free (XF/SF) condition; hDPSC products were subsequently stored by two-step banking including a master cell bank (MCB) and a working cell bank (WCB). The final products were directly thawed hDPSCs from the WCB. We tested the safety and quality check, stem cell properties, and preclinical potentials of final hDPSC products and hDPSC products in the MCB and WCB. Results: We optimized manufacturing procedures to isolate and expand hDPSC products under a XF/SF culture condition and established the MCB and the WCB. The final hDPSC products and hDPSC products in the MCB and WCB were validated the safety and quality including population doubling ability, chromosome stability, microorganism safety, and stem cell properties including morphology, cell surface marker expression, and multipotency. We also evaluated the in vivo immunogenicity and tumorigenicity and validated in vivo therapeutic efficacy for liver regeneration in a CCl 4-induced chronic liver fibrosis mouse model in the final hDPSC products and hDPSC products in the WCB.

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“…In majority of clinical trials involving MSCs published to date, the cells are used at early passages (no more than passage 5). However, cells with clonal aneuploidy and translocations have been described already at 1 st and 4 th passages of various MSC lines [ 3 , 4 , 5 ]. An extensive body of data suggests that MSCs are relatively genetically stable during culturing in vitro [ 6 – 9 , 10 , 11 ].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2018

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Background aimsSpontaneous 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.MethodsThe 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.ResultsThe 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.ConclusionsThe 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.

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Methodological Guidelines for Genetic Safety Testing of Cell Transplants

Cited by 9 publications

References 16 publications

Genotoxicity of Single-Walled Carbon Nanotubes: In Vitro Study on Human Embryonic Fibroblast Cells

Genotoxicity of Single-Walled Carbon Nanotubes: In Vitro Study on Human Embryonic Fibroblast Cells

A model study for the manufacture and validation of clinical-grade deciduous dental pulp stem cells for chronic liver fibrosis treatment

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

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