Genomic instability in long-term culture of human adipose-derived mesenchymal stromal cells

Abstract

Mesenchymal stromal/stem cells stem (MSC) have been widely studied due to their great potential for application in tissue engineering and regenerative and translational medicine. In MSC-based therapy for human diseases, cell proliferation is required to obtain a large and adequate number of cells to ensure therapeutic efficacy. During in vitro culture, cells are under an artificial environment and manipulative stress that can affect genetic stability. Several regulatory agencies have established guidelines to ensure greater safety in cell-based regenerative and translational medicine, but there is no specific definition about the maximum number of passages that ensure the lowest possible risk in MSC-based regenerative medicine. In this context, the aim of this study was to analyze DNA damage and chromosome alterations in adipose-derived mesenchymal stromal cells (ADMSC) until the eleventh passage and to provide additional subsidies to regulatory agencies related to number of passages in these cells. Thus, two methods in genetic toxicology were adopted: comet assay and micronucleus test. The comet assay results showed an increase in DNA damage from the fifth passage onwards. The micronucleus test showed a statistically significant increase of micronucleus from the seventh passage onwards, indicating a possible mutagenic effect associated with the increase in the number of passages. Based on these results, it is important to emphasize the need to assess genetic toxicology and inclusion of new guidelines by regulatory agencies to guarantee the safety of MSC-based therapies for human diseases.

Efficacy of adipose‐derived mesenchymal stem cell therapy in the treatment of chronic micro‐ and macrovascular complications of diabetes

“…140 There is an association between genomic instability (DNA damage and chromosome changes) and long cell culture or high passage numbers, which should be taken into account in cell therapies to guarantee the highest possible safety. 141 The increase in cell immunogenicity after MSC transplantation may constitute another concern for clinical applications.…”

DNA Methylation Profiles Are Stable in H3 K27M-Mutant Diffuse Midline Glioma Neurosphere Cell Lines

“…The clonal evolution of somatic TP53 mutations and MDM2 amplification have been observed in newly created chondrosarcoma cell lines grown as tumor spheres [6]. A temporal analysis of an adipose-derived mesenchymal stromal cell line highlighted genetic divergence, but only with the fifth passage, demonstrating the culture stability of DNA in the first few passages [7]. One study showed the stability of gene expression and copy number profiles in adult glioblastoma cell lines [8].…”