Generation of iPSC line from urine cells of hemophilia A with F8 (p. R814X) mutation

• Published in: Stem cell research 2022-04, Vol.60, p.102682-102682, Article 102682
• Main Authors: Sun, Wenwen, Hu, Xuchen, Wang, Lingyu, Ma, Yanchun, Zhang, Xialin, Zhang, Ruijuan, Zhao, Lidong, Ren, Juan, Yang, Linhua, Wang, Gang
• Format: Article
• Language: English
• Subjects: Cell Differentiation; Factor VIII - genetics; Hemophilia A - genetics; Hemophilia A - therapy; Humans; Induced Pluripotent Stem Cells; Mutation - genetics
• ISSN: 1873-5061; 1876-7753
• DOI: 10.1016/j.scr.2022.102682

•Expressing OSKM plasmid are not integrated into the cell genome.•We generated hiPSC of hemophilia A with F8(p.R814X) mutation from urine cell.•The iPSC models could be used to investigate Hemophilia mechanism. The lack of coagulation factor VIII in patient with nonsense mutation hemophilia A leads to varying degrees of bleeding symptoms, and long-term use of alternative therapies can produce inhibitors that affect the efficacy. In this study, human induced pluripotent stem cells (iPSCs) of hemophilia A were generated by reprogramming of urine cells. Human urine cells (HUCs) were isolated by collecting patients' mid-stream urine, and cultured to good state in urine medium. Then, the HUCs were transfected with PEP4-EO2S-ET2K and pCEP4-M2L, and iPSCs were obtained in the medium without trophoblast cells and the composition was determined. Finally, alkaline phosphatase staining, karyotype analysis, immunofluorescence staining and teratoma were used to verify that we successfully reprogrammed hemophilia A-specific human induced pluripotent stem cells from patients' urine cells, providing a safe and effective cell model for the study of molecular mechanism and related treatment of hemophilia.