J. Anim Sci.
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J. Anim Sci. 1978. 46:1241-1249.
© 1978 American Society of Animal Science
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Clonal Analysis of Satellite Cells in Growing Mice1

Ronald B. Young2 ,3 ,4,,5, Timothy R. Miller2 and Robert A. Merkel2 ,4,

Michigan State University, East Lansing 48824

5 To whom all correspondence should be addressed: Department of Food Science and Human Nutrition, Michigan State University, East Lansing, 48824.

Abstract

Viable, mononucleated cells were liberated from minced hind limb skeletal muscle of mice by incubation with .1% pronase at 37 C for 1 hour. Mice immediately after birth, at the most rapid growth stage and near maximum body weight were utilized as the source of skeletal muscle. Cell suspensions were placed in cell culture under conditions that permitted evaluation of several general properties of the cell population. The total number of viable cells/ gram of muscle decreased sharply during growth to a minimum value of approximately 2 x 104 cells/gram. The composition of this total cell population was evaluated in cell culture by cloning the cells at low density. Each colony of cells originating from a single cell was evaluated microscopically after 3 weeks in culture to determine if it was myogenic as evidenced by the presence of multinucleated myotubes. The percentage of myogenic clones was 26.2%, 23.7% and 17.0% for neonatal, rapidly growing and young adult mice muscle, respectively. Minimum values for the quantity of satellite cells/gram of muscle and the number of satellite cells/hind limb were calculated at each of the three ages. Both the concentration of satellite cells and the total quantity of satellite cells in hind limb muscle declined during growth. The combination of cell culture and cloning is useful for obtaining a measure of changes in viable satellite cell number under different conditions; however, a number of limitations are inherent and are emphasized.

Footnotes

1 Michigan Agricultural Experiment Station Journal Article No. 8298. This research was supported in part by funds from the Muscular Dystrophy Association of America, Michigan Agricultural Experiment Station Project Numbers 1241 and 1265H, and Biomedical Research Support Grants from the College of Agriculture and Natural Resources and the College of Osteopathic Medicine. Assistance with some of the cell culture experiments by Teresa Phillips is gratefully acknowledged.

2 Department of Animal Husbandry.

3 Department of Biomechanics.

4 Department of Food Science and Human Nutrition.






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Copyright © 1978 by the American Society of Animal Science.