Integration-free induced pluripotent stem cells derived from a patient with autosomal recessive Alport syndrome (ARAS)

Bernd Kuebler; Begoña Aran; Laia Miquel-Serra; Yolanda Muñoz; Elisabet Ars; Gemma Bullich; Monica Furlano; Roser Torra; Merce Marti; Anna Veiga; Angel Raya

doi:10.1016/j.scr.2017.08.021

Integration-free induced pluripotent stem cells derived from a patient with autosomal recessive Alport syndrome (ARAS)

Stem Cell Res. 2017 Dec:25:1-5. doi: 10.1016/j.scr.2017.08.021. Epub 2017 Sep 14.

Authors

Affiliations

¹ Center of Regenerative Medicine in Barcelona (CMRB), Barcelona, Spain; National Stem Cell Bank-Barcelona Node, Biomolecular and Bioinformatics Resources Platform PRB2, ISCIII, CMRB, Barcelona, Spain.
² Center of Regenerative Medicine in Barcelona (CMRB), Barcelona, Spain.
³ Molecular Biology Laboratory, Fundacio Puigvert, Instituto de Investigaciones Biomedicas Sant Pau (IIB-Sant Pau), Universitat Autonoma de Barcelona, REDinREN, Instituto de Investigacion Carlos III, Barcelona, Spain.
⁴ Inherited Kidney Disorders, Nephrology Department, Fundacio Puigvert, Instituto de Investigaciones Biomedicas Sant Pau (IIB-Sant Pau), Universitat Autonoma de Barcelona, REDinREN, Instituto de Investigacion Carlos III, Barcelona, Spain.
⁵ Center of Regenerative Medicine in Barcelona (CMRB), Barcelona, Spain; National Stem Cell Bank-Barcelona Node, Biomolecular and Bioinformatics Resources Platform PRB2, ISCIII, CMRB, Barcelona, Spain; Center for Networked Biomedical Research on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain.
⁶ Center of Regenerative Medicine in Barcelona (CMRB), Barcelona, Spain; Center for Networked Biomedical Research on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain; Institució Catalana de Recerca I Estudis Avançats (ICREA), Barcelona, Spain. Electronic address: [email protected].

PMID: 29246570
DOI: 10.1016/j.scr.2017.08.021

Abstract

A skin biopsy was obtained from a 25-year-old female patient with autosomal recessive Alport syndrome (ARAS) with the homozygous COL4A3 mutation c.345delG, p.(P166Lfs*37). Dermal fibroblasts were derived and reprogrammed by nucleofection with episomal plasmids carrying OCT3/4, SOX2, KLF4 LIN28, L-MYC and p53shRNA. The generated induced Pluripotent Stem Cell (iPSC) clone AS FiPS1 Ep6F-2 was free of genomically integrated reprogramming genes, had the specific homozygous mutation, a stable karyotype, expressed pluripotency markers and generated embryoid bodies which were differentiated towards the three germ layers in vitro. This iPSC line offers a useful resource to study Alport syndrome pathomechanisms and drug testing.

MeSH terms

Adult
Cells, Cultured
Cellular Reprogramming / genetics
Cellular Reprogramming / physiology
Exons / genetics
Female
Fibroblasts / cytology
Fibroblasts / metabolism
Humans
In Vitro Techniques
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / metabolism*
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors / genetics
Kruppel-Like Transcription Factors / metabolism
Mutation / genetics
Nephritis, Hereditary / metabolism*
Octamer Transcription Factor-3 / genetics
Octamer Transcription Factor-3 / metabolism
Organic Cation Transport Proteins / genetics
Organic Cation Transport Proteins / metabolism
Plasmids / genetics
Proto-Oncogene Proteins c-myc / genetics
Proto-Oncogene Proteins c-myc / metabolism
RNA-Binding Proteins / genetics
RNA-Binding Proteins / metabolism
SOXB1 Transcription Factors / genetics
SOXB1 Transcription Factors / metabolism
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism

Substances

KLF4 protein, human
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors
Lin28A protein, human
MYCL protein, human
Octamer Transcription Factor-3
Organic Cation Transport Proteins
POU5F1 protein, human
Proto-Oncogene Proteins c-myc
RNA-Binding Proteins
SOXB1 Transcription Factors
Tumor Suppressor Protein p53
solute carrier family 22 (organic cation transporter), member 3