A Pilot Analysis of Whole Transcriptome of Human Cryopreserved Sperm

Int J Mol Sci. 2024 Apr 8;25(7):4131. doi: 10.3390/ijms25074131.


Sperm cryopreservation is a procedure widely used to store gametes for later use, to preserve fertility in patients prior to gonadotoxic treatments or surgery, and for sperm donation programs. The purpose of the study was to assess the impact of cryopreservation on human sperm transcriptome. Semen samples were collected from 13 normospermic men. Each sample was divided into two aliquots. The total RNA was immediately extracted from one aliquot. The second aliquot was frozen and total RNA was extracted after a week of storage in liquid nitrogen. The RNA samples were randomized in four pools, each of six donors, and analyzed by microarrays. The paired Significance Analysis of Microarray was performed. We found 219 lower abundant transcripts and 28 higher abundant transcripts in cryopreserved sperm than fresh sperm. The gene ontology analysis disclosed that cryopreservation alters transcripts of pathways important for fertility (i.e., spermatogenesis, sperm motility, mitochondria function, fertilization, calcium homeostasis, cell differentiation, and early embryo development), although the increase of some transcripts involved in immune response can compensate for the harmful effects of freezing.

Keywords: fertility preservation; human sperm; microarray; sperm; sperm cryopreservation; sperm donation; transcriptome.

MeSH terms

  • Cryopreservation
  • Humans
  • Male
  • RNA
  • Semen*
  • Sperm Motility / genetics
  • Spermatozoa
  • Transcriptome*


  • RNA