Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope

L. Nel-Themaat, M. C. Gómez, P. Damiani, G. Wirtu, B. L. Dresser, K. R. Bondioli, Leslie A Lyons, C. E. Pope, R. A. Godke

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16 Scopus citations

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°C/min; 14%) ram semen, whereas no attachment occurred in the fast-frozen (10°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.

Original languageEnglish (US)
Pages (from-to)576-584
Number of pages9
JournalReproduction, Fertility and Development
Volume19
Issue number4
DOIs
StatePublished - 2007

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Keywords

  • Cell culture
  • Cryopreservation
  • Ejaculate
  • Epithelial
  • Gulf Coast native sheep
  • Nuclear transfer

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Reproductive Medicine
  • Developmental Biology
  • Endocrinology

Cite this

Nel-Themaat, L., Gómez, M. C., Damiani, P., Wirtu, G., Dresser, B. L., Bondioli, K. R., Lyons, L. A., Pope, C. E., & Godke, R. A. (2007). Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope. Reproduction, Fertility and Development, 19(4), 576-584. https://doi.org/10.1071/RD06153