Document Detail

Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope.
MedLine Citation:
PMID:  17524303     Owner:  NLM     Status:  MEDLINE    
Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh eland (Taurotragus oryx) semen. Cells attached to the culture surface from fresh (29%), cooled (43%) and slow-frozen (1 degrees C/min; 14%) ram semen, whereas no attachment occurred in the fast-frozen (10 degrees C/min) group. Proliferation occurred in fresh (50%) and cooled (100%) groups, but no cells proliferated after passage 1 (P1). Eland semen yielded cell lines (100%) that were cryopreserved at P1. In samples from GCN and cross-bred milk, cell attachment (83% and 95%, respectively) and proliferation (60% and 37%, respectively) were observed. Immunocytochemical detection of cytokeratin indicated an epithelial origin of semen-derived cells, whereas milk yielded either fibroblasts, epithelial or a mixture of cell types. Deoxyribonucleic acid microsatellite analysis using cattle-derived markers confirmed that eland cells were from the semen donor. Eland epithelial cells were transferred into eland oocytes and 12 (71%), six (35%) and two (12%) embryos cleaved and developed to morulae or blastocyst stages, respectively. In conclusion, we have developed a technique for obtaining somatic cells from semen. We have also demonstrated that semen-derived cells can serve as karyoplast donors for nuclear transfer.
L Nel-Themaat; M C Gómez; P Damiani; G Wirtu; B L Dresser; K R Bondioli; L A Lyons; C E Pope; R A Godke
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Reproduction, fertility, and development     Volume:  19     ISSN:  1031-3613     ISO Abbreviation:  Reprod. Fertil. Dev.     Publication Date:  2007  
Date Detail:
Created Date:  2007-05-25     Completed Date:  2007-08-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8907465     Medline TA:  Reprod Fertil Dev     Country:  Australia    
Other Details:
Languages:  eng     Pagination:  576-84     Citation Subset:  IM    
Department of Animal Sciences, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
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MeSH Terms
Biological Markers / analysis
Cell Culture Techniques
Cell Proliferation
DNA / analysis
Embryo Transfer
Extinction, Biological
Milk / chemistry,  cytology*
Nuclear Transfer Techniques
Semen / chemistry,  cytology*
Sheep, Domestic*
Specimen Handling / methods*
Reg. No./Substance:
0/Biological Markers; 9007-49-2/DNA

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