The goal of this project is to use tissue culture techniques to introduce upper-level biology students to current concepts in molecular cell biology. A sterile hood CO2 incubator, inverted and epifluoresence microscopes, a cryo-storage thermos, and a vertical slab gel electrophoresis system were purchased to implement these experiments. Students isolate membrane fractions from cultured cells using ultracentrifugation techniques. Membranes and whole cells are stained using primary antibodies to specific proteins and organelles and peroxidase-labelled and fluorescent-labelled secondary antibodies. Viral transformation is demonstrated using primary chick fibroblasts and Rous sarcoma virus, and students observe and quantitate the transformation utilizing the inverted microscope and pulse-labelling techniques. Chemical signaling is demonstrated through the addition of defined hormones to a BC3H1 cell line that cause differentiation of these cells into the muscle phenotype. Western and Northern blotting are also demonstrated with tissue-cultured cells.
