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OPTIMIZING GOAT SKIN FIBROBLAST CULTURE CONDITIONS FOR CLONING

Abstract

To maintain the nuclear integrity of somatic cells that provide beneficial traits for cloning, tissue culture techniques are used to culture cells in vitro using a growth medium supplemented with serum. Scientists have commonly used growth medium containing 10% serum when growing cells. The aim of this project was to examine the effect of different Fetal Bovine Serum (FBS) concentrations from 0-70% added to growth medium on goat skin fibroblast proliferation. 20,000 fibroblast cells were added to 1.5 ml of DMEM growth medium supplemented with 0, 10, 20, 30, 40, 50, 60 or 70 percent FBS in each of the wells of a 24-well microtiter plate. Cells were grown for three days at 37 degrees Celsius in a CO2 incubator. Results showed that addition of FBS to the medium increased the proliferation of fibroblast cells where maximal proliferation was observed at a concentration of 50% FBS. Further increase in FBS concentration to 60% and 70% led to a decline in fibroblast proliferation. The growth curve obtained using 10% FBS showed a decline in cell numbers after nine days. In contrast, cells grown in 50% FBS showed consistent growth until day twelve with a 15-fold increase in cell count as compared to cells grown in 10% FBS. In conclusion, to obtain an adequate number of cells for cloning, scientists can more efficiently grow fibroblast cells using 50% FBS which allows for a significant increase in cell proliferation as compared to the existing method of using 10% FBS.

Acknowledgements

Dr. Mahipal Singh, FVSU Department of Animal Science

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