Generation of patient-specific pluripotent induced stem cell line UFRJi007-A from a Brazilian familial amyotrophic lateral sclerosis patient

Gubert, Fernanda; Vasques, Juliana Ferreira; Cozendey, Tatiane D.; Domizi, Pablo; Toledo, María Fernanda; Kasai-Brunswick, Taís Hanae; Hochman‐Mendez, Camila; Campos, Mário; Zembrzuski, Verônica Marques; Loureiro, Marli P.S.; Lima, J.A. De; Gress, Claudio H.; Cabello, Giselda M. K.; Cabello, Pedro Hernán; Borgonovo, Tamara; Vaz, Isadora May; Silva, Rosane; Mendez-Otero, Rosália

doi:10.1016/j.scr.2019.101490

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“…Expression of pluripotency markers was determined by RT-PCR and flow cytometry (BD Accuri C6, BD FACSAria II and FlowJo software version 1). The reprogramming method used has already been validated ( Kasai-Brunswick et al, 2018 ; Mesquita et al, 2019 ; Gubert et al, 2019a ; Gubert et al ., 2019b ).…”

Section: Methodsmentioning

confidence: 99%

Chromosomal aberrations after induced pluripotent stem cells reprogramming

et al. 2021

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Induced pluripotent stem cells (iPSCs) are generated from adult cells that have been reprogrammed to pluripotency. However, in vitro cultivation and genetic reprogramming increase genetic instability, which could result in chromosomal abnormalities. Maintenance of genetic stability after reprogramming is required for possible experimental and clinical applications. The aim of this study was to analyze chromosomal alterations by using the G-banding karyotyping method applied to 97 samples from 38 iPSC cell lines generated from peripheral blood or Wharton’s jelly. Samples from patients with long QT syndrome, Jervell and Lange-Nielsen syndrome and amyotrophic lateral sclerosis and from normal individuals revealed the following chromosomal alterations: acentric fragments, chromosomal fusions, premature centromere divisions, double minutes, radial figures, ring chromosomes, polyploidies, inversions and trisomies. An analysis of two samples generated from Wharton’s jelly before and after reprogramming showed that abnormal clones can emerge or be selected and generate an altered lineage. IPSC lines may show clonal and nonclonal chromosomal aberrations in several passages (from P6 to P34), but these aberrations are more common in later passages. Many important chromosomal aberrations were detected, showing that G-banding is very useful for evaluating genetic instability with important repercussions for the application of iPSC lines.

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