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Implementation of an in-house flow cytometric analysis of DNA fragmentation in spermatozoa

Asian Journal of Andrology 2020年第3期22卷 246-251页

作者： Anne Sofie Rex Chunsen Wu jorn aagaard Jens Fedder Centre of Andrology and Fertility Clinic Odense University HospitalOdense 5000Denmark aagaard Gynaecological Clinic Skejby 8200AarhusDenmark Research Unit of Gynaecology and Obstetrics Faculty of HealthUniversity of Southern DenmarkOdense 5000Denmark.

An increased amount of DNA fragme ntation in the spermatozoa(SDF)is linked to male in fertility.The Sperm Chromati n Structure Assay(SCSA)is widely used for analysis of SDF.However,the current software(SCSASoftR)linked to this assay is licensed and often located within larger diagnostic centers.In this study,we present a protocol for using other types of software than SCSASoftR to determine the SDF index(DFI)with clinical relevance.This protocol is engineered after collect!ng and analyzing 254 samples from fertility patients and sperm donors over a 15-month period.DFI is analyzed using a strict protocol where the spermatozoa are treated with a strong acid(pH 1.2)followed by acridine orange.DFI is determined by a standard flow cytometric software,FACSDiva 6.1.3.Analysis of the outcome of the fertility treatment is included for 137 patients receiving either intrauterine inseminations(IUI)or timed coitus(TC).The results show that the chance of pregnancy decli nes as DFI in creases.We also found that the male DFI affects the chanee of pregnancy independent of the female age.We have shown that a standard flow cytometric software can be used when determi ning a clinical releva nt DFI.These findings are a sign ificant step toward impleme nting the an alysis as a part of the routi ne,in・house diag no sing of the male fertility patient and subseque ntly optimizing the treatme nt course of the couple with reduced human and financial costs.

关键词： DNA fragme ntation flow cytometry in-house an alysis in trauteri ne in semi natio n Sperm Chromati n Structure Assay spermatozoa

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