title: Induced pluripotent stem cells from patients with focal cortical dysplasia and refractory epilepsy.
abstract: Focal cortical dysplasia (FCD) is caused by numerous alterations, which can be divided into abnormalities of the cortical architecture and cytological variations; however, the exact etiology of FCD remains unknown. The generation of induced pluripotent stem cells (iPSCs) from the cells of patients with neurological diseases, and their subsequent tissue‑specific differentiation, serves as an invaluable source for testing and studying the initial development and subsequent progression of diseases associated with the central nervous system. A total of 2 patients demonstrating seizures refractory to drug treatment, characterized as FCD Type IIb, were enrolled in the present study. Fibroblasts were isolated from residual skin fragments obtained from surgical treatment and from brain samples obtained during surgical resection. iPSCs were generated following exposure of fibroblasts to viral vectors containing POU class 5 homeobox 1 (OCT4), sex determining region Y‑box 2 (SOX2), Kruppel‑like factor 4 and c‑MYC genes, and were characterized by immunohistochemical staining for the pluripotent markers homeobox protein NANOG, SOX2, OCT4, TRA1‑60 and TRA1‑81. The brain samples were tested with antibodies against protein kinase B (AKT), phosphorylated‑AKT, mechanistic target of rapamycin (mTOR) and phosphorylated‑mTOR. Analysis of the AKT/mTOR pathway revealed a statistically significant difference between the cerebral tissues of the two patients, which were of different ages (45 and 12 years old). Clones with the morphological features of embryonic cells were detected on the 13th day and were characterized following three subcultures. The positive staining characteristics of the embryonic cells confirmed the successful generation of iPSCs derived from the patients' fibroblasts. Therefore, the present study presents a method to obtain a useful cellular source that may help to understand embryonic brain development associated with FCD.
authors.affiliation: Brain Institute of Rio Grande do Sul (BraIns), Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Daniel Rodrigo
authors.initials: DR
authors.lastname: Marinowic
authors.affiliation: Institute for Biomedical Research, Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Fernanda
authors.initials: F
authors.lastname: Majolo
authors.affiliation: Brain Institute of Rio Grande do Sul (BraIns), Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Alessandra Deise
authors.initials: AD
authors.lastname: Sebben
authors.affiliation: Laboratory of Pathological Anatomy, Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Vinicius Duval
authors.initials: VD
authors.lastname: da Silva
authors.affiliation: Laboratory of Pathological Anatomy, Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Tiago Giuliani
authors.initials: TG
authors.lastname: Lopes
authors.affiliation: Epilepsy Surgery Program, Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Eliseu
authors.initials: E
authors.lastname: Paglioli
authors.affiliation: Postgraduate Program in Medicine and Health Sciences, Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: André
authors.initials: A
authors.lastname: Palmini
authors.affiliation: Brain Institute of Rio Grande do Sul (BraIns), Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Denise Cantarelli
authors.initials: DC
authors.lastname: Machado
authors.affiliation: Brain Institute of Rio Grande do Sul (BraIns), Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, RS 90610000, Brazil.
authors.forename: Jaderson Costa
authors.initials: JC
authors.lastname: da Costa
mesh_terms: D002478:Cells, Cultured; D065150:Cellular Reprogramming; D002648:Child; D004827:Epilepsy / Q000378:metabolism / Q000473:pathology*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000473:pathology*; D008297:Male; D065703:Malformations of Cortical Development, Group I / Q000378:metabolism / Q000473:pathology*; D008875:Middle Aged; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism
chemical_list: C546842:MTOR protein, human; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases