title: Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope.
abstract: 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.
authors.affiliation: Department of Animal Sciences, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
authors.forename: L
authors.initials: L
authors.lastname: Nel-Themaat
authors.forename: M C
authors.initials: MC
authors.lastname: Gómez
authors.forename: P
authors.initials: P
authors.lastname: Damiani
authors.forename: G
authors.initials: G
authors.lastname: Wirtu
authors.forename: B L
authors.initials: BL
authors.lastname: Dresser
authors.forename: K R
authors.initials: KR
authors.lastname: Bondioli
authors.forename: L A
authors.initials: LA
authors.lastname: Lyons
authors.forename: C E
authors.initials: CE
authors.lastname: Pope
authors.forename: R A
authors.initials: RA
authors.lastname: Godke
mesh_terms: D000818:Animals; D000866:Antelopes; D015415:Biomarkers / Q000032:analysis; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D015925:Cryopreservation; D004247:DNA / Q000032:analysis; D004624:Embryo Transfer; D053476:Extinction, Biological; D005260:Female; D008297:Male; D008892:Milk / Q000737:chemistry / Q000166:cytology*; D053652:Nuclear Transfer Techniques; D012661:Semen / Q000737:chemistry / Q000166:cytology*; D034561:Sheep, Domestic; D013048:Specimen Handling / Q000379:methods*
chemical_list: D015415:Biomarkers; D004247:DNA