| Flow cytometric sorting of frozen-thawed spermatozoa in sheep and non-human primates. | |
| | |
MedLine Citation:
|
PMID: 14984693 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Research was conducted in sheep to determine an effective preparation method for high-purity sorting of frozen-thawed spermatozoa. The efficacy of sorting frozen-thawed spermatozoa was then investigated in several non-human primate species. An aliquot of each ejaculate (three rams, three ejaculates per ram) was processed as a fresh control (FRESH). Frozen spermatozoa were thawed and prepared for sorting by no further processing (FT-NEAT), washing (FT-WASH) or gradient centrifugation (FT-GRADIENT) and evaluated for motility at 1 h post-staining and motility and acrosomal status at 0 and 4 h post-sorting. Samples were analysed using a high-speed cell sorter. High levels of purity for X- and Y-enriched samples were achieved for all treatments (85-92%). The percentage of motile spermatozoa before sorting was lower (P < 0.05) for frozen-thawed samples (FT-NEAT: 32.7 +/- 2.5%; FT-WASH: 32.2 +/- 3.3%; FT-GRADIENT: 73.9 +/- 3.7%) compared with FRESH (83.3 +/- 1.2%). Post-sorting, the percentage of motile spermatozoa before and after incubation for FT-NEAT (60.0 +/- 5.1% and 27.2 +/- 6.1% for 0 and 4 h, respectively) was lower than that for FRESH (87.8 +/- 0.9% and 83.3 +/- 1.2% for 0 and 4 h, respectively; P < 0.05), FT-WASH (80.0 +/- 2.4% and 71.7 +/- 3.6% for 0 and 4 h, respectively; P < 0.05) and FT-GRADIENT (84.4 +/- 1.3% and 77.2 +/- 1.7% for 0 and 4 h, respectively; P < 0.05). Vanguard sperm migration distance through artificial cervical mucus was lower (P < 0.05) for FT-NEAT (17.7 +/- 1.7 mm) compared with FT-WASH (29.1 +/- 3.8 mm) and FT-GRADIENT (28.4 +/- 2.0 mm) and similar (P < 0.05) to FRESH (23.7 +/- 1.8 mm). Sample preparation using a modified wash method enabled high-purity sorting (range 86-97% purity) of frozen-thawed epididymal spermatozoa in the baboon (Papio hamadryas), common marmoset (Callithrix jacchus) and common chimpanzee (Pan troglodytes). For all non-human primate species, sorted spermatozoa were progressively motile (marmoset: 20.5 +/- 5.5%; baboon: 37.5 +/- 2.5%; chimpanzee: 73.0 +/- 2.0%), acrosome intact (marmoset: 68.5 +/- 7.5%; baboon: 89.5 +/- 1.5%; chimpanzee: 84.0 +/- 1.0%) and able to penetrate an artificial cervical mucus. In summary, high-purity sorting of frozen-thawed ram and non-human primate spermatozoa with recovery of progressively motile, acrosome-intact spermatozoa was possible after processing to remove cryodiluent. |
| | |
Authors:
|
J K O'Brien; F K Hollinshead; K M Evans; G Evans; W M C Maxwell |
Related Documents
:
|
21444563 - Medical facilities and encounters at the 2010 vancouver winter olympic games. 14556353 - Medical care for people under detention. 18829713 - Antenatal bartter's syndrome: why is this not a lethal condition? |
Publication Detail:
|
Type: Journal Article |
Journal Detail:
|
Title: Reproduction, fertility, and development Volume: 15 ISSN: 1031-3613 ISO Abbreviation: Reprod. Fertil. Dev. Publication Date: 2003 |
Date Detail:
|
Created Date: 2004-02-26 Completed Date: 2007-09-24 Revised Date: 2009-01-16 |
Medline Journal Info:
|
Nlm Unique ID: 8907465 Medline TA: Reprod Fertil Dev Country: Australia |
Other Details:
|
Languages: eng Pagination: 367-75 Citation Subset: IM |
Affiliation:
|
Centre for Advanced Technologies in Animal Genetics and Reproduction, Faculty of Veterinary Science, The University of Sydney, NSW, Australia. justineo@vetsci.usyd.edu.au |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Cryopreservation* Flow Cytometry Male Primates* Semen Preservation* Sex Preselection / methods* Sheep, Domestic* Spermatozoa / physiology* |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Birth of a cloned calf derived from a vitrified hand-made cloned embryo.
Next Document: Improved sperm cryopreservation using cold cryoprotectant.