title: Reprogramming of mouse renal tubular epithelial cells to induced pluripotent stem cells.
abstract: Kidney disease has reached epidemic proportions and is associated with high mortality and morbidity rates. Stem cell-based therapy may effectively treat kidney damage by cell transplantation. The breakthrough discovery using a combination of four transcription factors to reprogram genetically somatic cells into induced pluripotent stem (iPS) cells was a milestone in stem cell therapy. The lentivirus was packaged containing OCT4, SOX2, c-MYC and KLF4 transcription factors and then transfected mouse renal tubular epithelial cells (RTECs). The colonies were picked up at 21 days and were tested by cytochemistry, immunofluorescence assay and quantitative real-time polymerase chain reaction. Viral transgene expression levels were also assessed by quantitative analysis. Additionally, embryoid bodies from iPS cells were formed, and immunofluorescence and teratoma assays were performed. Karyotype analysis of mouse RTEC-derived iPS cells was also performed. The iPS cells were indistinguishable from mouse embryonic stem cells with respect to colony morphology, the expression of pluripotency-associated transcription factors and surface markers, embryoid body-mediated differentiation potential and teratoma assays. Quantitative polymerase chain reaction demonstrated that the lentiviral transgenes were largely silenced. The mouse RTEC-derived iPS cells exhibited a normal karyotype of 40,XY. iPS cells can be produced using mouse RTECs, which would be helpful in investigations to ameliorate the symptoms of kidney disease and to slow the progression of kidney disease by in vitro and in vivo animal studies.
authors: Department of Nephrology, Jimin Hospital, Shanghai, PR China.
authors: Wei-Wei
authors: WW
authors: Wang
authors: Wei
authors: W
authors: Wang
authors: Yan
authors: Y
authors: Jiang
authors: Guo-Feng
authors: GF
authors: Han
authors: Shi
authors: S
authors: Lu
authors: Gangqiang
authors: G
authors: Li
authors: Jinyuan
authors: J
authors: Zhang
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D064987:Cell- and Tissue-Based Therapy; D065150:Cellular Reprogramming; D053595:Embryonic Stem Cells / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007684:Kidney Tubules / Q000166:cytology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D016086:Lentivirus; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics; D014162:Transfection
chemical_list: C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse