Generation of integration-free induced pluripotent stem cell lines derived from two patients with X-linked Alport syndrome (XLAS)

Stem Cell Res. 2017 Dec:25:291-295. doi: 10.1016/j.scr.2017.08.019. Epub 2017 Sep 9.

Abstract

Skin biopsies were obtained from two male patients with X-linked Alport syndrome (XLAS) with hemizygous COL4A5 mutations in exon 41 or exon 46. Dermal fibroblasts were extracted and reprogrammed by nucleofection with episomal plasmids carrying OCT3/4, SOX2, KLF4 LIN28, L-MYC and p53 shRNA. The generated induced Pluripotent Stem Cell (iPSC) lines AS-FiPS2-Ep6F-28 and AS-FiPS3-Ep6F-9 were free of genomically integrated reprogramming genes, had the specific mutations, a stable karyotype, expressed pluripotency markers and generated embryoid bodies which were differentiated towards the three germ layers in vitro. These iPSC lines offer a useful resource to study Alport syndrome pathomechanisms and drug testing.

MeSH terms

  • Adult
  • Cell Line
  • Cells, Cultured
  • Cellular Reprogramming
  • Collagen Type IV / genetics
  • Collagen Type IV / metabolism
  • Embryoid Bodies / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Karyotype
  • Kruppel-Like Factor 4
  • Male
  • Mutation
  • Nephritis, Hereditary / genetics*
  • Nephritis, Hereditary / metabolism

Substances

  • COL4A5 protein, human
  • Collagen Type IV
  • KLF4 protein, human
  • Kruppel-Like Factor 4