Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection

The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen–thawed rhesus monkey spermatozoa and fertili...

Full description

Saved in:
Bibliographic Details
Published in:Theriogenology 2005-06, Vol.63 (9), p.2356-2371
Main Authors: Yeoman, Richard R., Mitalipov, Shoukhrat, Gerami-Naini, Behzad, Nusser, Kevin D., Wolf, Don P.
Format: Article
Language:English
Subjects:
animal genetic resources
Animals
Cryopreservation
Cryopreservation - methods
Cryopreservation - veterinary
Embryo Transfer - veterinary
Female
Fertility
Fertilization
freezing
germplasm conservation
Hot Temperature
ICSI
Macaca mulatta
Male
male fertility
Pregnancy
Rhesus monkey
Semen Preservation - methods
Semen Preservation - veterinary
Sperm Injections, Intracytoplasmic - veterinary
Sperm Motility
Spermatozoa
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen–thawed rhesus monkey spermatozoa and fertility via intracytoplasmic spermatozoa injection (ICSI) of sibling oocytes. Washed spermatozoa were frozen in straws or in pellets using different cryoprotective media and processed post-thaw with or without a density gradient centrifugation step. Among the four study series, motility post-thaw was improved with density gradient centrifugation (17–24% versus 75%, P < 0.01) achieving levels similar to fresh spermatozoa. Spermatozoa injected oocytes (total n = 377) were co-cultured on BRL cells and observed for fertilization and development. With spermatozoa frozen in straws in liquid nitrogen vapors, the fertilization rate after ICSI was lower than with fresh spermatozoa (40–44% versus 77–86%, P < 0.05), even with the Percoll-enriched fraction that exhibited robust motility. In contrast, somewhat slower freezing of spermatozoa in pellets on dry ice supported fertilization rates (73%) that were similar to the fresh counterpart. Developmental rates of fertilized eggs were similar in all experiments. A total of 106 embryo transfers has resulted in the first primate born after ICSI with F/T ejaculated spermatozoa plus 22 other infants to date. Additionally, a 3–4 h incubation after thawing improved the fertilization rate with spermatozoa from a male with poor post-thaw recovery of sperm motility. In conclusion, an acceptable fertilization rate after ICSI with motile, frozen–thawed primate spermatozoa was observed comparable to that obtained with fresh spermatozoa allowing small quantities of competent spermatozoa to be used with ICSI to facilitate propagation of desirable primate genotypes.
ISSN:0093-691X
1879-3231
DOI:10.1016/j.theriogenology.2004.05.033