title: Molecular karyotyping of single sperm with nuclear vacuoles identifies more chromosomal abnormalities in patients with testiculopathy than fertile controls: implications for ICSI.
abstract: OBJECTIVE
  Is there a difference between molecular karyotype of single sperm selected by high-magnification microscopy from infertile patients with testicular damage and from proven fertile controls?
  
  
  CONCLUSIONS
  The molecular karyotype of single sperm from patients with testiculopathy had a significantly higher percentage of chromosomal alterations than fertile controls.
  
  
  BACKGROUND
  Infertile patients with testicular impairment have many sperm with aneuploidies and/or increased structural chromosome alterations. In these patients, sperm use by ICSI has poor outcome and raises concerns about the possible impact on pregnancy loss and transmission of genes abnormalities in offspring. High-magnification microscopy has been recently introduced to select morphologically better sperm aimed at improving ICSI outcome. However, there are no studies evaluating the molecular karyotype of sperm selected by this method.
  
  
  METHODS
  Three consecutive infertile patients with oligozoospermia due to testicular damage and three age-matched proven fertile men attending a tertiary care center, were enrolled in the study from September to November 2014. Inclusion criteria of patients were age ≥30 ≤35 years, at least 2 years of infertility, oligozoospermia (sperm count below 10 million), reduced testicular volumes high FSH plasma levels and absence of altered karyotype, Y chromosome microdeletions, cystic fibrosis transmembrane conductance regulator gene mutations, sperm infections, cigarette smoking, varicocele, obesity.
  
  
  METHODS
  Participants were evaluated for sperm parameters, sex hormones and testicular color-doppler ultrasound. From each semen sample, 20 sperm with large vacuoles (LVs), 20 with small vacuoles (SVs) and 20 with no vacuoles (NVs) were retrieved individually by a micromanipulator system. Each cell was further analyzed by whole genome amplification and array comparative genomic hybridization (aCGH).
  
  
  RESULTS
  The aCGH allowed us to detect chromosomal aneuploidies, unbalanced translocations and complex abnormalities. Sperm selected from infertile patients showed a higher percentage of abnormal molecular karyotypes than controls (19.4 versus 7.7%, respectively, P < 0.001). In particular, sperm with LV and SV showed 38.3 and 20.0% abnormal karyotype in infertile men versus 18.3 and 5.0% in controls, respectively (both P < 0.01). Complex abnormalities were found only in the LV category. An abnormal karyotype was never found in NV sperm from both patients and controls.
  
  
  CONCLUSIONS
  The main limitation of this study is the low number of included subjects. Moreover, a time of writing we have no data regarding the ICSI outcome using LV, SV or NV sperm. This is the first study evaluating the molecular karyotype of single sperm selected by high-magnification microscopy and further confirmation of the data is needed.
  
  
  CONCLUSIONS
  Our data showed that sperm from infertile patients with testicular impairment have a higher percentage of abnormal molecular karyotypes than sperm from fertile controls. Therefore, if confirmed, our data suggest that the use of individually retrieved NV sperm may improve ICSI outcome in infertile men with testicular damage.
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Garolla
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Sartini
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Cosci
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Damiano
authors.initials: D
authors.lastname: Pizzol
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Ghezzi
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bertoldo
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Menegazzo
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Speltra
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ferlin
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Via Giustiniani 2, Padova 35121, Italy carlo.foresta@unipd.it.
authors.forename: Carlo
authors.initials: C
authors.lastname: Foresta
mesh_terms: D000328:Adult; D002869:Chromosome Aberrations; D006801:Humans; D007621:Karyotyping / Q000379:methods*; D008297:Male; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa; D013733:Testicular Diseases / Q000473:pathology*; D014617:Vacuoles
keywords: array comparative genomic hybridization; high-magnification microscopy; male infertility; molecular karyotype; oligozoospermia; sperm vacuoles; whole genome amplification