Assessment of three generations of mice derived by ICSI using freeze-dried sperm

Ming Wen Li, Brandon J. Willis, Stephen M Griffey, Jimmy L. Spearow, Kevin C K Lloyd

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


Although the derivation of mice by intracytoplasmic sperm injection (ICSI) using freeze-dried sperm has been demonstrated previously, a comprehensive analysis of their viability, health, and fertility has not. The purpose of the present study was to determine the extent to which ICSI using freeze-dried sperm stored at 4 C for 12 months from mice on either an inbred (C57BL/6J) or hybrid (B6D2F1/J) genetic background results in genomic instability and/or phenotypic abnormality in mice and two generations of their progeny. Fertilization rates (number of 2-cells per injected oocytes) using ICSI of fresh and freeze-dried sperm were similar within and between mouse strains, although fewer freeze-dried sperm-derived embryos than fresh sperm-derived embryos developed to blastocysts in vitro (C57BL/6J and B6D2F1/J) and liveborn pups in vivo (B6D2F1/J only). Nevertheless, once born, mice derived by ICSI using freeze-dried sperm in both mouse strains were healthy and reproductively sound. No major differences in litter size, weaning rate, and sex ratio were noted in the two generations of progeny (F2 and F3) of ICSI-derived offspring using freeze-dried sperm compared with that in the natural mating (control) group. Further, there was no evidence that either ICSI or freeze drying induced genomic instability, as determined by microsatellite analysis of the derived mice and subsequent generations when compared with both parental genotypes, nor were there differences in the number or types of pathological changes in any of the three generations of progeny. We conclude that viable, healthy and genomically stable mice can be derived by ICSI using freeze-dried mouse sperm stored in the refrigerator for at least 2 months. Further, because freeze drying is a simpler and more economical technique compared with embryo and sperm cryopreservation, the results of this study justify additional research to continue to develop and enhance the technique for the preservation, storage, and sharing of genetically altered mice.

Original languageEnglish (US)
Pages (from-to)239-251
Number of pages13
Issue number3
StatePublished - Aug 2009


  • Freeze drying
  • ICSI
  • Microsatellite
  • Mouse
  • Sperm

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology


Dive into the research topics of 'Assessment of three generations of mice derived by ICSI using freeze-dried sperm'. Together they form a unique fingerprint.

Cite this