Genomic alterations in human umbilical cord–derived mesenchymal stromal cells call for stringent quality control before any possible therapeutic approach

Borghesi, A.; Avanzini, Maria Antonietta; Novara, Francesca; Mantelli, Melissa; Lenta, Elisa; Achille, Valentina; Cerbo, Rosa Maria; Tzialla, Chryssoula; Longo, Stefania; Silvestri, Annalisa De; Zimmermann, Luc J. I.; Manzoni, Paolo; Zecca, Marco; Spinillo, Arsenio; Maccario, Rita; Zuffardi, Orsetta; Stronati, Mauro

doi:10.1016/j.jcyt.2013.06.006

Cited by 23 publications

(17 citation statements)

References 35 publications

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“…Our findings are in line with previous reports suggesting that BM-MSCs may occasionally develop genetic aberrations after long-term culture, without evidence of malignant transformation whatsoever [35–37]. As regards WJ-MSCs, contradictory results have been reported so far for the genomic stability of UC-derived MSCs [38–42]. The inconsistencies are probably related to the variability of MSC populations, culture conditions and methodologies (classical karyotype, FISH or comparative genomic hybridization array).…”

Section: Discussionsupporting

confidence: 92%

Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of Wharton’s jelly and bone marrow-derived mesenchymal stem cells

et al. 2017

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BackgroundIn view of the current interest in exploring the clinical use of mesenchymal stem cells (MSCs) from different sources, we performed a side-by-side comparison of the biological properties of MSCs isolated from the Wharton’s jelly (WJ), the most abundant MSC source in umbilical cord, with bone marrow (BM)-MSCs, the most extensively studied MSC population.MethodsMSCs were isolated and expanded from BM aspirates of hematologically healthy donors (n = 18) and from the WJ of full-term neonates (n = 18). We evaluated, in parallel experiments, the MSC immunophenotypic, survival and senescence characteristics as well as their proliferative potential and cell cycle distribution. We also assessed the expression of genes associated with the WNT- and cell cycle-signaling pathway and we performed karyotypic analysis through passages to evaluate the MSC genomic stability. The hematopoiesis-supporting capacity of MSCs from both sources was investigated by evaluating the clonogenic cells in the non-adherent fraction of MSC co-cultures with BM or umbilical cord blood-derived CD34+ cells and by measuring the hematopoietic cytokines levels in MSC culture supernatants. Finally, we evaluated the ability of MSCs to differentiate into adipocytes and osteocytes and the effect of the WNT-associated molecules WISP-1 and sFRP4 on the differentiation potential of WJ-MSCs.ResultsBoth ex vivo-expanded MSC populations showed similar morphologic, immunophenotypic, survival and senescence characteristics and acquired genomic alterations at low frequency during passages. WJ-MSCs exhibited higher proliferative potential, possibly due to upregulation of genes that stimulate cell proliferation along with downregulation of genes related to cell cycle inhibition. WJ-MSCs displayed inferior lineage priming and differentiation capacity toward osteocytes and adipocytes, compared to BM-MSCs. This finding was associated with differential expression of molecules related to WNT signaling, including WISP1 and sFRP4, the respective role of which in the differentiation potential of WJ-MSCs was specifically investigated. Interestingly, treatment of WJ-MSCs with recombinant human WISP1 or sFRP4 resulted in induction of osteogenesis and adipogenesis, respectively. WJ-MSCs exhibited inferior hematopoiesis-supporting potential probably due to reduced production of stromal cell-Derived Factor-1α, compared to BM-MSCs.ConclusionsOverall, these data are anticipated to contribute to the better characterization of WJ-MSCs and BM-MSCs for potential clinical applications.

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Section: Discussionsupporting

confidence: 92%

Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of Wharton’s jelly and bone marrow-derived mesenchymal stem cells

et al. 2017

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“…At least 20 metaphases should be analysed, and in the presence of aberrations, 20 more metaphases are requested, as defined by the General Guidelines and Quality Assurance for Cytogenetics [47,[55][56]. The karyotyping analysis would be sufficient as a release test with the exclusion of two identical abnormal metaphases on 20 This article has been peer-reviewed and accepted for publication, but has yet to undergo copyediting and proof correction.…”

Section: Karyotypementioning

confidence: 99%

Ex Vivo Expanded Mesenchymal Stromal Cell Minimal Quality Requirements for Clinical Application

Torre

Lucarelli

GuidiSimona³

et al. 2015

Stem Cells and Development

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Mesenchymal Stromal Cells (MSC), as advanced therapy products, must satisfy all the requirements for human use of medicinal products, aiming to maintain the quality and safety of the cells. The MSC manufacturing process for clinical use should comply with the principles of Good Manufacturing Practice (GMP). This ensures that cell preparations are produced and controlled, from the collection and manipulation of raw materials, through the processing of intermediate products, to the quality controls, storage, labelling and packaging, and release.The objective of this document is to provide the minimal quality requirements for the MSC production and its delivery for clinical use, so the safety of the final cell therapy product will not be compromised. For this purpose, the document evaluates the most important steps of GMP-compliant MSC production: the isolation and expansion process; the validation phase of the process, including all quality controls for the characterization, functionality, potency and safety of MSCs; the quality control at the batch release to guarantee the safety of patient infusion. This opinion paper reflects the consensus viewpoint of the authors and scientists participating the GISM Working Group*.

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“…возникновению раз-личных структурных и численных хромосомных аномалий. При культивировании клеток в отдаленных пассажах в клеточных культурах могут возникать новые клоны с хромосомными перестройками, которые могут характеризоваться более высо-кими темпами пролиферации и лучшей выживаемостью после криоконсервации, чем исходные клетки с нормальным гено-мом [5, 33,34]. При нестабильности генома (главным образом, под влиянием условий культивирования) наряду с клональ-ными перестройками хромосом появляются неклональные, которые создают генетическое разнообразие в популяции, что обеспечивает ее выживание и адаптацию к неблагоприятным условиям [35].…”

Section: причины генетической нестабильностиunclassified

Use of Cytogenetic Analysis Methods for Assessing the Quality of Cell Lines in Biomedical Cell Products

Рачинская¹,

Меркулов²

2018

Biopreparaty. Profilaktika, diagnostika, lechenie

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Биомедицинские клеточные продукты (БМКП) -новая группа препаратов, основанных на применении клеточных линий различного происхождения для лечения широкого спектра заболеваний, в том числе в сфере регенеративной медицины. Контроль качества клеточного компонента таких препаратов явля-ется важной задачей на всех стадиях разработки и производства БМКП. Большое внимание должно уде-ляться подтверждению безопасности препаратов в силу ряда их особенностей и возможности возникно-вения побочных эффектов при их применении, в том числе риска развития онкологических заболеваний. Возможной причиной канцерогенеза может стать генетическая нестабильность клеточного компонента БМКП. Для выявления генетической нестабильности клеток, входящих в состав БМКП, на хромосом-ном уровне возможно применение ряда цитогенетических методов. Подтверждение наличия в клетках неизменного кариотипа и идентификацию различных хромосомных аномалий возможно осуществлять с помощью как классических цитогенетических методов анализа, например дифференциальное окраши-вание хромосом, так и с помощью молекулярно-цитогенетических методов, основанных на применении флуоресцентной гибридизации in situ. При комплексном использовании этих методов возможно получе-ние достоверной оценки генетической стабильности и косвенного доказательства отсутствия малигниза-ции клеточной линии в составе БМКП. Biomedical cell products (BMCPs) are a new group of biologicals that are based on various cell lines and are used in the treatment of a wide range of diseases as well as in the field of regenerative medicine. The quality control of the cellular component in such preparations is very important at all stages of BMCPs development and production. Much attention should be given to confirmation of BMCPs safety because of their specific properties and potential side effects, including the risk of cancer development. Carcinogenesis may be attributed to genetic instability of the BMCP cellular component. A number of cytogenetic methods can be used at the chromosomal level in order to identify the genetic instability of cells in a BMCP. Confirmation of the normal karyotype of cells and identification of various chromosomal abnormalities can be achieved using both classic cytogenetic analysis methods, such as chromosome banding, and molecular cytogenetic methods based on the use of fluorescent in situ hybridization. Combination of these methods may provide a reliable estimation of genetic stability of the cell line in a BMCP, and indirect evidence of absence of malignancy.

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Genomic alterations in human umbilical cord–derived mesenchymal stromal cells call for stringent quality control before any possible therapeutic approach

Cited by 23 publications

References 35 publications

Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of Wharton’s jelly and bone marrow-derived mesenchymal stem cells

Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of Wharton’s jelly and bone marrow-derived mesenchymal stem cells

Ex Vivo Expanded Mesenchymal Stromal Cell Minimal Quality Requirements for Clinical Application

Use of Cytogenetic Analysis Methods for Assessing the Quality of Cell Lines in Biomedical Cell Products

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