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 urinederived cryptorchid-specific induced pluripotent stem cells could present a good human genetic system for future studies investigating the molecular mechanism of cryptorchidism.

Urine-derived stem cells: A novel and versatile progenitor source for cell-based therapy and regenerative medicine

“…The latter characteristic of urine cells contributes to improved reprogramming efficiency. Our data and other investigators' studies 66 , 67 , 68 , 69 , 70 demonstrate the feasibility of rapid and efficient iPS cell generation from a sample of human urine, suggesting potential therapeutic uses of patient- derived iPS cells both for the treatment of various genetic defects via cell-based therapy and for drug screening.…”

Generating a Non-Integrating Human Induced Pluripotent Stem Cell Bank from Urine-Derived Cells

“…However, these approaches remain inefficient or laborious. We and others had previously described that epithelia-like cells could be isolated from human urine and could be further reprogrammed into iPS cells with high efficiency by using virus-based approach [13], [14], [15], [16]. Human UCs can be conveniently obtained through non-invasive method, thus representing an ideal cell source for iPS cell generation.…”

Urine-derived induced pluripotent stem cells as a modeling tool to study rare human diseases

“…Cryptorchidism involves multiple causes, such as genetic mutations and environmental factors (65,66). Researchers have established cryptorchid-specific iPSC lines with genetic variations and they have found that urine cells may represent a better source for generation of iPSCs, especially in the study of pediatric diseases (60). Most patients with complex disorders are treated with various medications over a prolonged period, which may have an impact on donor cell propagation and nuclear reprogramming.…”

“…Because iPSCs resemble ESCs in terms of pluripotency and the potential for self-renewal, patient-specific iPSCs could represent a promising strategy for investigating the molecular mechanisms of disease and discovery of drugs to treat them (93,94). Recent studies have reported that generating UiPSCs from patients with rare diseases would provide insight into the mechanisms of those diseases (59)(60)(61)(62)(63)(64). The availability of limitless amounts of diseased cells from patients with a rare disease could, along with gene editing, prove useful in clinical practice (Figure 1).…”