Study question In men with Raised Sperm DNA Fragmentation Index (DFI), Would advance sperm preparation techniques at Intra-Cytoplasmic Sperm Injection (ICSI) alter embryo euploidy status? Summary answer In men with raised Sperm DFI, use of advance sperm selection techniques are safe and do not appear to alter ploidy status of the embryos What is known already MACS (Magnetic Activated Cell Sorting), Microfluidic Sperm Sorting (MF), surgically retrieved Testicular Sperm (TESA) and Double Density Gradient sperm processing (DDG) have been shown to optimize sperm selection at ICSI for men with raised sperm DFI. Would any of these sperm selection methods alter embryo ploidy status is not proven Study design, size, duration This was a retrospective observational study conducted at a private fertility centre in-between years 2015 & 2022. Couples where women (<37yrs) had idiopathic Recurrent Implantation Failures (RIF) and men had raised sperm DFI (>25% DFI) and underwent Pre-Implantation Genetic Testing (PGT) were evaluated (n = 473). Based on sperm selection method the study population was divided into four groups. TESA (n = 84), MACS (n = 105), MF(n = 135), DDG (n = 148).A total of 1802 embryos were evaluated in this study. Participants/materials, setting, methods All women underwent Controlled Ovarian Stimulation (COS) and oocyte retrieval and male partners underwent sperm processing as per the clinics SOP. Appropriate counselling and written consents were obtained from all couples, where applicable. All oocytes subjected to ICSI, extended blastocyst culture and trophoectoderm (TE) biopsy. Post biopsy embryos were vitrified and TE tissue subjected to Next-Generation Sequencing (NGS) to assess the ploidy status. Ploidy status between various sperm selection methods was compared. Main results and the role of chance Euploidy rates between the study groups were as follows: TESA (n = 311 embryos screened)- 48% MACS (n = 445 embryos screened) – 51% MF (n = 489 embryos screened) – 61% DDG (n = 557 embryos screened)- 54% Though numerically the ploidy status was highest in MF group for euploid embryos, there seems to be no statistical significance between the groups (p-value = 0.973). Different sperm selection techniques do not seem to alter embryo ploidy status. Ejaculated sperm or Testicular sperms had similar euploidy status. Use of Testicular sperm, which is considered immature doesn’t seem to increase embryo aneuploidy. MACS, MF and DDG all seemed to have similar euploid embryos. This further warrants research to validate the role of MACS and MF as a routine intervention to optimize sperm selection and reproductive outcomes. Limitations, reasons for caution Retrospective design, un-equal group size and small sample size, a well-designed study is needed to further validate the findings of this study. Wider implications of the findings Role of advance sperm selection methods to optimize embryo implantation is yet to be proven. Data from this study showed no alarming increase in embryo aneuploidy in younger women and was comparable with published literature. Use of advance sperm selection techniques seems safe and doesn’t alter embryo ploidy status. Trial registration number not applicable

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