title: Generation and characterization of human cryptorchid-specific induced pluripotent stem cells from urine.
abstract: Cryptorchidism is a common congenital birth defect in human beings with the possible complication of infertility. An in vitro model of cryptorchidism might be valuable due to the inaccessibility of human embryos for research purposes. In this study, we reprogrammed urine cells containing genetic variations in insulin-like factor 3, zinc finger (ZNF) 214, and ZNF215 from a cryptorchid patient by introducing human OCT4, SOX2, C-MYC, and KLF4 with lentivirus. The cells were then replated on irradiated mouse embryonic fibroblasts and cultured with the human embryonic stem (ES) cell medium. The compact colonies with well-defined borders were manually picked, and 2 induced pluripotent cell lines were fully characterized. Our results demonstrated that these 2 cell lines were similar to human ES cells in morphological appearance, marker expression, and epigenetic status of the pluripotent cell-specific gene, OCT4. These cells could be differentiated into cells of all 3 germ layers in teratomas and in vitro, including into the VASA-positive germ cell lineage. Both parental urine cells and the reprogrammed cells possessed the normal karyotype and the same short tandem repeat loci, indicating that these 2 cell population share the same genetic identity. This establishment and characterization of human urine-derived cryptorchid-specific induced pluripotent stem cells could present a good human genetic system for future studies investigating the molecular mechanism of cryptorchidism.
authors.affiliation: Central Laboratory, Children's Hospital of Shanghai, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Junmei
authors.initials: J
authors.lastname: Zhou
authors.forename: Xue
authors.initials: X
authors.lastname: Wang
authors.forename: Shengli
authors.initials: S
authors.lastname: Zhang
authors.forename: Yijun
authors.initials: Y
authors.lastname: Gu
authors.forename: Ling
authors.initials: L
authors.lastname: Yu
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.forename: Tongbin
authors.initials: T
authors.lastname: Gao
authors.forename: Fang
authors.initials: F
authors.lastname: Chen
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D003456:Cryptorchidism / Q000378:metabolism / Q000473:pathology*; D004268:DNA-Binding Proteins / Q000235:genetics; D014644:Genetic Variation; D005822:Genetic Vectors; D005855:Germ Layers; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D007328:Insulin / Q000235:genetics*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D016086:Lentivirus / Q000235:genetics; D008297:Male; D009154:Mutation; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D011506:Proteins / Q000235:genetics*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics; D014556:Urine / Q000166:cytology*
chemical_list: D004268:DNA-Binding Proteins; D007328:Insulin; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C084797:Leydig insulin-like protein; C489427:MYC protein, human; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D011506:Proteins; D016271:Proto-Oncogene Proteins c-myc; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C409745:ZNF214 protein, human; C409744:ZNF215 protein, human