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United States Department of Agriculture

Agricultural Research Service

Title: Evaluation of Primary Rumen Epithelial Cell Incubation Techniques in Sheep

Authors
item Gillis, R - UNIVERSITY OF TENNESSEE
item Klotz, J - UNIVERSITY OF TENNESSEE
item Baldwin, Ransom
item Heitmann, R - UNIVERSITY OF TENNESSEE

Submitted to: Small Ruminant Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 26, 2002
Publication Date: August 1, 2002
Citation: GILLIS, R.C., KLOTZ, J.L., BALDWIN, R.L., HEITMANN, R.N. EVALUATION OF PRIMARY RUMEN EPITHELIAL CELL INCUBATION TECHNIQUES IN SHEEP. SMALL RUMINANT RESEARCH. 2002.

Interpretive Summary: Objectives of this study were to determine if the number of cells incubated in primary rumen epithelial cell cultures affects production rates of metabolites and to standardize reporting criteria by obtaining an optimum mode of data expression. Rumen Epithelial cells were isolated from 5 crossbred sheep and were incubated in media containing volatile fatty acids. Different amounts of cells were included in order to assess the impact of cell number on incubation characteristics that might affect scientific interpretation. Metabolite production was determined and expressed as a function of cell number, dry matter or total protein alone or per epithelial wet tissue weight, body weight (BW) or metabolic BW to generate twelve different forms of data expression. A suggested range of rumen epithelial cells to include in incubations is 5 to 20-million/flask. This range will minimize the potential for altered metabolite production caused by incubating large cell quantities as well as the experimental error associated with using low cell numbers. When rumen tissue is taken from animals of the same species, size and stage of development, data adjusted by cell number is preferred.

Technical Abstract: Objectives of this study were to determine if the number of cells incubated in primary rumen epithelial cell cultures affects production rates of metabolites and to standardize reporting criteria by obtaining an optimum mode of data expression. Rumen Epithelium was excised from 5 crossbred sheep and cells were isolated. Isolated cells were incubated in 25 mM propionate and 10 mM butyrate at concentrations of .5, 1, 5, 10, 20 and 40 million cells per flask. Production of acetoacetate (ACAC), - hydroxybutyrate ( HBA), lactate (LAC) and pyruvate (PYR) were determined and expressed as a function of cell number, dry matter or total protein alone or per epithelial wet tissue weight, body weight (BW) or metabolic BW to generate twelve different forms of data expression. Data expressed per cells resulted in the lowest variation (P < .05) and data adjusted for metabolic BW had less variation than BW. ACAC concentrations were largest at .5-million cells/flask (P < .05) and there were not different between 1 to 20 while 40 differed from .5 and 5-million cells/flask. HBA concentrations were largest at 1 and 5-million cells/flask. However, 1 and 5 only differed (P < .05) from 20 and 40-million cells/flask. HBA:ACAC ratios were below one for .5 and 1-million cells/flask indicating low mitochondrial redox potentials (P < .05). A suggested range of rumen epithelial cells to include in incubations is 5 to 20-million/flask. This range will minimize the potential for altered metabolite production caused by incubating large cell quantities as well as the experimental error associated with using low cell numbers. When rumen tissue is taken from animals of the same species, size and stage of development, data adjusted by cell number is preferred.

Last Modified: 12/22/2014
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