关键词: 梗阻性无精子症, 非梗阻性无精子症, 诱导性多能干细胞, 仙台病毒

Abstract: Objective To establish testis-derived induced pluripotent stem cell (iPSCs) of patients with obstructive azoospermia (OA) and non-obstructive azoospermia (NOA), and contrast the difference in the two lines process. Methods 5 to 6 mm3 testicular tissue of 3 OA and idiopathic NOA patients were excised for primary culture, transfected Sendai virus in 3rd or 4th passages, and transferred cell on feeder after they are dense. Then pick the iPS clone on the 21st to the 22nd day, establish iPSCs line and identify iPSCs pluripotency and differentiation potential. Results Two iPSCs lines were successfully established, there were no significant differences in the two lines. Both AP stains were positive, Octamer-binding transcription factor 4 (OCT4), SRY-related-box protein-2 (SOX2), and stage-specific embryonic antigen- 4 (SSEA-4) and tumor rejection antigen-1-60 (TRA-1-60) were all positive for immunofluorescence staining; teratoma test in mice also showed that the obtained two iPSCs could differentiate to three-dimensional germ layers. Conclusion Using the Sendai virus non-genomic integration transfection approach, testis-derived iPSCs can be established in OA and idiopathic NOA patients.

Key words: Obstructive azoospermia, Non-obstructive azoospermia, Induced pluripotent stem cells, Sendai virus