Identification of structural DNA variations in human cell cultures after long-term passage

Abstract

Random amplified polymorphic DNA (RAPD) analysis was adapted for genomic identification of cell cultures and evaluation of DNA stability in cells of different origin at different culture passages. DNA stability was observed in cultures after no more than 5 passages. Adipose-derived stromal cells demonstrated increased DNA instability. RAPD fragments from different cell lines after different number of passages were cloned and sequenced. The chromosomal localization of these fragments was identified and single-nucleotide variations in RAPD fragments isolated from cell lines after 8-12 passages were revealed. Some of them had permanent localization, while most variations demonstrated random distribution and can be considered as de novo mutations.

A novel transposable element based authentication protocol for Drosophila cell lines

“…2013). Apart from gross genomic changes, extensive passaging introduced single nucleotide polymorphisms in mammalian cell lines (P avlova et al . 2015).…”

“…Extensive passaging of cell lines can potentially alter cellular genomes (WENGER et al 2004;OH et al 2013). Apart from gross genomic changes, extensive passaging introduced single nucleotide polymorphisms in mammalian cell lines (PAVLOVA et al 2015). To determine the effect of extensive passaging on the gTED signatures generated in this study, we passaged S2R+ cell line 50 times and isolated genomic DNA in triplicate at every tenth passage for processing (Fig.…”

“…2013). Likewise, long term passaging of mammalian cell lines is known to lead to increased single nucleotide variations (P avlova et al . 2015), reduced differentiation potential (Y ang et al .…”

A novel transposable element-based authentication protocol for Drosophila cell lines

“…For example, extensive passaging (>50 passages) of viral-transformed human lymphoblastoid cell lines is associated with increased genotypic instability ( Oh et al 2013 ). Likewise, long-term passaging of mammalian cell lines is known to lead to increased single nucleotide variations ( Pavlova et al 2015 ), reduced differentiation potential ( Yang et al 2018 ), and changes in the karyotype ( Wenger et al 2004 ). To overcome these inconsistencies in experiments across laboratories when using human cell lines, the American National Standards Institute and the American Type Culture Collection (ANSI/ATCC ASN-002) have provided a standard for vertebrate cell culture work.…”

Head and neck cancer organoids established by modification of the CTOS method can be used to predict in vivo drug sensitivity

“…Establishment of cell lines from 2D cultures of tumors derived from patient tissue has a low success rate, and only select tumor cells can survive in the 2D environment which may be reflected in an altered tumor heterogeneity from the original tumor. Furthermore, it is well known that long term culture can select for genetic alterations [15–17]. Since most 3D culture models derived from established 2D cell lines do not maintain the original tumor phenotype, development of 3D cancer organoid cultures directly from a patient’s cancer tissue which includes cancer stem cells, can better recapitulate the character of the patient’s tumor than those derived from cells previously passaged as 2D cultures [18–21].…”