PMID- 15589288
OWN - NLM
STAT- MEDLINE
DCOM- 20050303
LR  - 20161124
IS  - 0093-691X (Print)
IS  - 0093-691X (Linking)
VI  - 63
IP  - 1
DP  - 2005 Jan 1
TI  - Flow cytometric sorting of non-human primate sperm nuclei.
PG  - 246-59
AB  - Pre-determination of the sex of offspring has implications for management and 
      conservation of captive wildlife species, particularly those with single 
      sex-dominated social structures. Our goal is to adapt flow cytometry technology 
      to sort spermatozoa of non-human primate species for use with assisted 
      reproductive technologies. The objectives of this study were to: (i) determine 
      the difference in DNA content between X- and Y-bearing spermatozoa (ii) sort 
      sperm nuclei into X- and Y-enriched samples; and (iii) assess the accuracy of 
      sorting. Spermatozoa were collected from two common marmosets (Callithrix 
      jacchus), seven hamadryas baboons (Papio hamadryas) and two common chimpanzees 
      (Pan troglodytes). Human spermatozoa from one male were used as a control. Sperm 
      nuclei were stained (Hoechst 33342), incubated and analyzed using a high-speed 
      cell sorter. Flow cytometric reanalysis of sorted samples (sort reanalysis, 
      10,000 events/sample) and fluorescence in situ hybridization (FISH; 500 sperm 
      nuclei/sample) were used to evaluate accuracy of sorting. Based on fluorescence 
      intensity of X- and Y-bearing sperm nuclei, the difference in DNA content between 
      X and Y populations was 4.09 +/- 0.03, 4.20 +/- 0.03, 3.30 +/- 0.01, and 2.97 +/- 
      0.05%, for marmoset, baboon, chimpanzee and human, respectively. Sort reanalysis 
      and FISH results were similar; combined data revealed high levels of purity for 
      X- and Y-enriched samples (94 +/- 0.9 and 93 +/- 0.8%, 94 +/- 0.7 and 94 +/- 
      0.5%, 91 +/- 0.9 and 97 +/- 0.6%, 94 +/- 0.6 and 94 +/- 0.9%, for marmoset, 
      baboon, chimpanzee and human, respectively). These data indicate the potential 
      for high-purity sorting of spermatozoa from non-human primates.
FAU - O'Brien, Justine K
AU  - O'Brien JK
AD  - Faculty of Veterinary Science, Centre for Advanced Technologies in Animal 
      Genetics and Reproduction, University of Sydney, NSW 2006, Australia. 
      justineo@vetsci.usyd.edu.au
FAU - Stojanov, Tomas
AU  - Stojanov T
FAU - Heffernan, Scott J
AU  - Heffernan SJ
FAU - Hollinshead, Fiona K
AU  - Hollinshead FK
FAU - Vogelnest, Larry
AU  - Vogelnest L
FAU - Maxwell, W M Chis
AU  - Maxwell WM
FAU - Evans, Gareth
AU  - Evans G
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Theriogenology
JT  - Theriogenology
JID - 0421510
RN  - 0 (Benzimidazoles)
RN  - 0 (Fluorescent Dyes)
RN  - 9007-49-2 (DNA)
RN  - P976261J69 (bisbenzimide ethoxide trihydrochloride)
SB  - IM
MH  - Animals
MH  - Benzimidazoles
MH  - Callithrix
MH  - Cell Nucleus/*chemistry
MH  - Cell Separation/methods/*veterinary
MH  - DNA/analysis
MH  - Flow Cytometry/methods/*veterinary
MH  - Fluorescent Dyes
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/veterinary
MH  - Male
MH  - Pan troglodytes
MH  - Papio hamadryas
MH  - *Primates
MH  - Sex Determination Analysis/*veterinary
MH  - Spermatozoa/*ultrastructure
MH  - X Chromosome
MH  - Y Chromosome
EDAT- 2004/12/14 09:00
MHDA- 2005/03/04 09:00
CRDT- 2004/12/14 09:00
PHST- 2004/03/09 00:00 [received]
PHST- 2004/04/21 00:00 [accepted]
PHST- 2004/12/14 09:00 [pubmed]
PHST- 2005/03/04 09:00 [medline]
PHST- 2004/12/14 09:00 [entrez]
AID - S0093691X04001475 [pii]
AID - 10.1016/j.theriogenology.2004.04.013 [doi]
PST - ppublish
SO  - Theriogenology. 2005 Jan 1;63(1):246-59. doi: 
      10.1016/j.theriogenology.2004.04.013.