Chromosomal scan of single sperm cells by combining fluorescence-activated cell sorting and next-generation sequencing

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http://hdl.handle.net/10138/298915

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Tran , Q T , Jatsenko , T , Poolamets , O , Tsuiko , O , Lubenets , D , Reimand , T , Punab , M , Peters , M & Salumets , A 2019 , ' Chromosomal scan of single sperm cells by combining fluorescence-activated cell sorting and next-generation sequencing ' Journal of Assisted Reproduction and Genetics , vol. 36 , no. 1 , pp. 91-97 . https://doi.org/10.1007/s10815-018-1340-0

Title: Chromosomal scan of single sperm cells by combining fluorescence-activated cell sorting and next-generation sequencing
Author: Tran, Quoc Ty; Jatsenko, Tatjana; Poolamets, Olev; Tsuiko, Olga; Lubenets, Dmitri; Reimand, Tiia; Punab, Margus; Peters, Maire; Salumets, Andres
Contributor: University of Helsinki, Clinicum
Date: 2019-01
Number of pages: 7
Belongs to series: Journal of Assisted Reproduction and Genetics
ISSN: 1058-0468
URI: http://hdl.handle.net/10138/298915
Abstract: PurposeThe purpose of this study was to develop a feasible approach for single sperm isolation and chromosome analysis by next-generation sequencing (NGS).MethodsSingle sperm cells were isolated from semen samples of normozoospermic male and an infertile reciprocal translocation (RcT) carrier with the 46,XY,t(7;13)(p12;q12.1) karyotype using the optimized fluorescence-activated cell sorting (FACS) technique. Genome profiling was performed using NGS.ResultsFollowing whole-genome amplification, NGS,and quality control, the final chromosome analysis was performed on 31 and 6 single cell samples derived from the RcT carrier and normozoospermic male, respectively. All sperm cells from normozoospermic male showed a normal haploid 23-chromosome profile. For the RcT carrier, the sequencing data revealed that 64.5% of sperm cells harbored different variants of chromosome aberrations, involving deletion of 7p or 7q, duplication of 7p, and duplication of 13q, which is concordant with the expected chromosome segregation patterns observed in balanced translocation carriers. In one sample, a duplication of 9q was also detected.ConclusionsWe optimized FACS protocol for simple and efficient isolation of single human sperm cells that subsequently enabled a successful genome-wide chromosome profiling and identification of segmental aneuploidies from these individual cells, following NGS analysis. This approach may be useful for analyzing semen samples of infertile men or chromosomal aberration carriers to facilitate the reproductive risk assessment.
Subject: Single sperm genomic analysis
Reciprocal translocation
Fluorescence-activated cell sorting
Whole-genome amplification
Next-generation sequencing
RECIPROCAL TRANSLOCATION CARRIERS
IN-SITU HYBRIDIZATION
ABNORMALITIES
SPERMATOZOA
ANEUPLOIDY
MEN
RECOMBINATION
RATES
MALES
3123 Gynaecology and paediatrics
1184 Genetics, developmental biology, physiology
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