In vitro separation of embryonic chick skeletal muscle myoblasts and fibroblasts: comparison of their characteristics.
The aim of the present experiments was to test two methods of separating myoblasts and fibroblasts (selective plating, differential trypsinization) from chick embryonal skeletal muscle and to compare their characteristics. Ornithine decarboxylase (ODC) activity, the amount of incorporated [3H]leucine into proteins and incorporation of [3H]thymidine into DNA were significantly higher in myoblasts than in fibroblasts separated by selective plating. When comparing myoblasts and fibroblasts separated by differential trypsinization, significantly higher ODC activity and greater incorporation of [3H]leucine into protein, but no incorporation of [3H]thymidine into DNA, were found in myoblasts. Higher ODC activity and greater incorporation of labelled leucine were found in fibroblasts separated by the selective plating than in fibroblasts separated by differential trypsinization. The incorporation of labelled thymidine into DNA was higher in myoblasts separated by selective plating than in myoblasts obtained by differential trypsinization. The method of selective plating appears to be simpler and adequate for obtaining myoblastic and fibroblastic cell cultures with sufficiently low mutual contamination. The method of differential trypsinization involves a more drastic treatment of cells and is more time consuming.
Lamosová
D
D
Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Ivanka pri Dunaji.
Juráni
M
M
Vaneková
M
M
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002642:Chick Embryo; D005347:Fibroblasts; D066298:In Vitro Techniques; D007930:Leucine / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D009955:Ornithine Decarboxylase / Q000378:metabolism*; D013936:Thymidine / Q000378:metabolism*
D009955:Ornithine Decarboxylase; D007930:Leucine; D013936:Thymidine