title: Generation of human iPSCs from cells of fibroblastic and epithelial origin by means of the oriP/EBNA-1 episomal reprogramming system.
abstract: BACKGROUND
  The prospect of therapeutic applications of the induced pluripotent stem cells (iPSCs) is based on their ability to generate virtually any cell type present in human body. Generation of iPSCs from somatic cells has opened up new possibilities to investigate stem cell biology, to better understand pathophysiology of human diseases, and to design new therapy approaches in the field of regenerative medicine. In this study, we focus on the ability of the episomal system, a non-viral and integration-free reprogramming method to derive iPSCs from somatic cells of various origin.
  
  
  METHODS
  Cells originating from neonatal and adult tissue, renal epithelium, and amniotic fluid were reprogrammed by using origin of replication/Epstein-Barr virus nuclear antigen-1 (oriP/EBNA-1)-based episomal vectors carrying defined factors. The iPSC colony formation was evaluated by using immunocytochemistry and alkaline phosphatase assay and by investigating gene expression profiles. The trilineage formation potential of generated pluripotent cells was assessed by embryoid body-mediated differentiation. The impact of additionally introduced factors on episome-based reprogramming was also investigated.
  
  
  RESULTS
  Reprogramming efficiencies were significantly higher for the epithelial cells compared with fibroblasts. The presence of additional factor miR 302/367 in episomal system enhanced reprogramming efficiencies in fibroblasts and epithelial cells, whereas the downregulation of Mbd3 expression increased iPSC colony-forming efficiency in fibroblasts solely.
  
  
  CONCLUSIONS
  In this study, we performed a side-by-side comparison of iPSC colony-forming efficiencies in fibroblasts and epithelial cells transiently transfected with episomal plasmids and demonstrated that iPSC generation efficiency was highest when donor samples were derived from epithelial cells. We determined that reprogramming efficiency of episomal system could be further improved. Considering results obtained in the course of this study, we believe that episomal reprogramming provides a simple, reproducible, and efficient tool for generating clinically relevant pluripotent cells.
authors.affiliation: Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland. anna.drozd@celther.com.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Drozd
authors.affiliation: Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland. maciej.walczak@celther.com.
authors.forename: Maciej P
authors.initials: MP
authors.lastname: Walczak
authors.affiliation: Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland. sylwester.piaskowski@celther.com.|Department of Tumor Biology, Medical University of Łódź, Żeligowskiego 7/9, 90-752, Łódź, Poland. sylwester.piaskowski@celther.com.
authors.forename: Sylwester
authors.initials: S
authors.lastname: Piaskowski
authors.affiliation: Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland. ewelina.stoczynska-fidelus@celther.com.|Department of Tumor Biology, Medical University of Łódź, Żeligowskiego 7/9, 90-752, Łódź, Poland. ewelina.stoczynska-fidelus@celther.com.
authors.forename: Ewelina
authors.initials: E
authors.lastname: Stoczynska-Fidelus
authors.affiliation: Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland. piotr.rieske@celther.com.|Department of Tumor Biology, Medical University of Łódź, Żeligowskiego 7/9, 90-752, Łódź, Poland. piotr.rieske@celther.com.
authors.forename: Piotr
authors.initials: P
authors.lastname: Rieske
authors.affiliation: Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland. dawid.grzela@celther.com.
authors.forename: Dawid P
authors.initials: DP
authors.lastname: Grzela
mesh_terms: D000650:Amnion / Q000166:cytology; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D052816:Foreskin / Q000166:cytology; D000066449:Human Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014556:Urine / Q000166:cytology
chemical_list: D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; C115369:MBD3 protein, human; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D014157:Transcription Factors; C417029:EBV-encoded nuclear antigen 1