Identifier

etd-04112005-195230

Degree

Master of Science (MS)

Department

Animal Science (Animal, Dairy, and Poultry Sciences)

Document Type

Thesis

Abstract

The influence of donor animal sex or age on in vitro bovine cell culture was evaluated to provide foundation information for the selection of donor tissue for nuclear transfer. Skin biopsies were taken from each of sixteen individuals including four bulls (B), four cows (C), four male calves (MC), four female calves (FC). At passage 2, cells from in vitro culture of cell lines were influenced not by gender but by age in the mean cell generation time (MGT). When evaluating familial lineage, comparison between related and unrelated groups showed that most comparisons do not show significant differences in lag time, stationary phase viable cell counts (SPCC) and MGT. In each cell line, there was high cell viability throughout the growth curves, indicating stable cell maintenance and proper cell harvest was conducted in this study. At passage 4, MGT of each cell line was not influenced by age but by sex at passage 2, however, at a later cell passage (by passage 4), the MGT of each cell line was not affected by either sex or age of the donor. By passage 4, the MGT of each cell line was not affected by either sex or age. As passages continued, the extrinsic environmental factors likely influenced the MGT. Cell cycle analysis at passage 4 on day 0 of this study showed that > 90% of cells were in G0/G1 portion in each cell line of all groups. Cell lines from younger donors were more frequently at higher G0/G1 percentages, or synchronized than those derived from older donors. Thus, age of donor animal could be a factor in selecting cell line for NT, especially when G0/G1 nuclei are intended for use. Male groups (B and MC) showed higher stationary phase viable cell counts than female groups (C and FC). Most comparisons showed no significant differences in lag time, SPCC and MGT between related and unrelated familial lineage groups. Each cell line showed constant viability (94.36 to 97.98%) at passage 4 throughout the growth curves.

Date

2005

Document Availability at the Time of Submission

Release the entire work immediately for access worldwide.

Committee Chair

Robert A. Godke

DOI

10.31390/gradschool_theses.846

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