Toward pre-conceptual genetic analysis of human spermatozoa

Dmitri Dozortsev, Rui Serafim, Jakson J. Cardoso, Soraya Abdelmassih, Peter Nagy, Michael P. Diamond, Roger Abdelmassih

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Nuclei of mature mammalian spermatozoa are extraordinarily resistant to chemical and thermal injury. Additionally, decondensation of spermatozoa DNA can be accompanied by little or no visual changes of the sperm head. This study tested whether human spermatozoa could be recovered following several cycles of primer extension preamplification (PEP) and used to achieve fertilization and subsequent development of human oocytes. An attempt was also made to amplify PEP buffer after spermatozoon removal. The results demonstrate that the sperm head can be successfully recovered following treatment with KOH or proteinase K followed by one to four cycles of PEP. It is also shown that following this treatment, the spermatozoa can be injected into the oocytes and will transform into a pronucleus if the oocyte is activated by sperm cytosolic fraction. In some cases, it was also possible to obtain polymerase chain reaction signals using a buffer after sperm cells were removed following several cycles of PEP. Although sperm participation in development was confirmed by fluorescence in-situ hybridization, light microscopy revealed some degree of damage to spermatozoal chromosomes. It is concluded that pre-conceptual analysis of sperm cells may be possible, but more research is necessary to determine the optimal conditions that would preserve sperm DNA integrity while allowing accurate diagnoses.

Original languageEnglish (US)
Pages (from-to)392-399
Number of pages8
JournalReproductive BioMedicine Online
Volume7
Issue number4
DOIs
StatePublished - Jan 1 2003
Externally publishedYes

Keywords

  • DNA
  • Oocyte
  • PGD
  • Polymerase chain reaction
  • Pronucleus
  • Spermatozoa

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology
  • Developmental Biology

Fingerprint Dive into the research topics of 'Toward pre-conceptual genetic analysis of human spermatozoa'. Together they form a unique fingerprint.

  • Cite this

    Dozortsev, D., Serafim, R., Cardoso, J. J., Abdelmassih, S., Nagy, P., Diamond, M. P., & Abdelmassih, R. (2003). Toward pre-conceptual genetic analysis of human spermatozoa. Reproductive BioMedicine Online, 7(4), 392-399. https://doi.org/10.1016/S1472-6483(10)61882-2