12 Jun You intend to grow human fibroblasts in culture
Question
You intend to grow human fibroblasts in culture to study their signal transduction pathways. Explain your answers to the following questions.
a) You obtain a sample of fibroblasts from a human volunteer and place them on a tissue culture dish. What might you include, or not include, in a serum-free culture for these cells? Why?
b) You wish to characterize these cells and determine what subunits of integrins that may be expressed.
Which integrin subtype would you predict to be the predominant class of integrin in these cells? Why? (which alpha and beta subunits?).
c) What ligand would you “coat” (incubate the culture dish with a solution of this protein ligand to allow association with the plastic) your tissue culture dishes with to provide your fibroblasts with the most likely ligand they interact with in vivo? Why?
d) Predict and explain what would happen if you reduced the calcium concentration in the media 100-fold.
e) Predict and explain what might happen if you included anti-RGD sequence antibodies in your fibroblast culture?
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