Generation of pluripotent stem cells using non-integrated approach in human azoospermia patients / 天津医药

ABSTRACT

Objective To establish induced pluripotent stem cells (iPSCs) in patients with azoospermia by non-integrated approach. Methods Using the commercially available serum-free medium (TeSR?2) and embryonic stem cell culture medium (Stem Adhere? Defined Matrix) to define the culture system, the iPSCs were established by using non-integrated Sendai virus infection in peripheral blood mononuclear cells of azoospermia patients. The immunofluorescence, karyotype analysis, embryoid body differentiation and teratoma formation were used to identify pluripotency, karyotype and differentiation ability of iPSCs. Results The established iPSCs showed the characteristics of human embryonic stem cells. Immunofluorescence analysis showed that octamer-binding transcription factor 4 (OCT4), SRY-related-box protein-2 (SOX2), stage-specific embryonic antigen-4 (SSEA-4) and tumor rejection antigen-1-60 (TRA-1-60) were positive for the expression of stem cell pluripotency markers. Karyotype analysis showed that they had normal karyotype. In addition, embryoid body and teratoma tests showed that the iPSCs had the ability to differentiate into three germ layers in vitro and in vivo. Conclusion The induction of pluripotent stem cell line is successfully constructed by non-integrated approach in azoospermia patients.