Academic Journal
Chromosomal scan of single sperm cells by combining fluorescence-activated cell sorting and next-generation sequencing
| العنوان: | Chromosomal scan of single sperm cells by combining fluorescence-activated cell sorting and next-generation sequencing |
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| المؤلفون: | Tran, Quoc Ty, Jatsenko, Tatjana, Poolamets, Olev, Tsuiko, Olga, Lubenets, Dmitri, Reimand, Tiia, Punab, Margus, Peters, Maire, Salumets, Andres |
| المساهمون: | Clinicum, Department of Obstetrics and Gynecology, University of Helsinki, HUS Gynecology and Obstetrics |
| بيانات النشر: | SPRINGER/PLENUM PUBLISHERS |
| سنة النشر: | 2019 |
| المجموعة: | Helsingfors Universitet: HELDA – Helsingin yliopiston digitaalinen arkisto |
| مصطلحات موضوعية: | 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, Gynaecology and paediatrics, Genetics, developmental biology, physiology |
| الوصف: | 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. ; Peer reviewed |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| ردمك: | 978-0-00-456212-4 0-00-456212-7 |
| Relation: | This study was funded by the Estonian Ministry of Education and Research (IUT34-16), Enterprise Estonia (EU48695), and the Horizon 2020 innovation program (WIDENLIFE, 692065).; 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; http://hdl.handle.net/10138/298915; 0ca4acc7-7cd0-43c0-8ed0-50802bcc4de1; 85056320854; 000456212700012 |
| الاتاحة: | http://hdl.handle.net/10138/298915 |
| Rights: | info:eu-repo/semantics/openAccess ; openAccess |
| رقم الانضمام: | edsbas.8924B879 |
| قاعدة البيانات: | BASE |
| ردمك: | 9780004562124 0004562127 |
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