Comparison of colony-formation efficiency of bovine fetal fibroblast cell lines cultured with low oxygen, hydrocortisone, L-carnosine, bFGF, or different levels of FBS.
A comparison of colony-formation efficiency (CFE) was made between six independent bovine fetal fibroblast (BFF) cell lines used in somatic cell nuclear transfer. Variation in CFE was assessed under different culture conditions. The conditions examined were ambient atmosphere (approximately 20% oxygen) culture versus 5% oxygen culture, three levels of fetal bovine serum (FBS) in the medium (5%, 10% or 20%), and the amendment of 10% FBS medium with basic fibroblast growth factor (1 ng/mL), L-carnosine (20 mM), or hydrocortisone (1 microM). The six BFF cell lines showed significant differences from one another in CFE. No significant difference in CFE was found with reduced oxygen culture. L-Carnosine also had no significant effect on CFE. A FBS concentration of 10% was found to produce the best overall CFE. Hydrocortisone treatment reduced the size of colonies although the number of colonies formed was not affected. Basic FGF increased the size of colonies but the number of colonies formed was not affected. The results showed that different BFF cell lines varied significantly in their CFE. Also, some medium supplements or culture conditions that have shown positive CFE effects on the fibroblasts of other species failed to show significant positive CFE effects on the BFF cell lines tested.
Talbot
Neil C
NC
USDA, ARS, Animal and Natural Resources Institute, Biotechnology and Germplasm Laboratory, Beltsville Agricultural Research Center, Maryland 20705, USA. Ntalbot@anri.barc.usda.gov
Powell
Anne M
AM
Caperna
Thomas J
TJ
D000818:Animals; D002336:Carnosine / Q000494:pharmacology; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D019976:Cloning, Organism / Q000379:methods*; D003114:Colony-Forming Units Assay; D003470:Culture Media; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects; D006854:Hydrocortisone / Q000494:pharmacology; D010100:Oxygen
D003470:Culture Media; D016222:Fibroblast Growth Factor 2; D002336:Carnosine; D010100:Oxygen; D006854:Hydrocortisone