麻省理工学院 生物材料课程 lecture 13 Quantifying Cell Behavior.pdf

3.051J/20.340J 1 Lecture 13: Quantifying Cell Behavior 1. Cell Cultures a. Why use cell cultures? in vivo (living organisms) in vitro (glass, i.e., culture) Pros 1. native 3D environment 1. simplified model systems 2. all relevant signals present 2. study parameters independently 3. observe as function of time Cons 1. many variables – noisy data 1. unnatural 2D environment 2. animal rights concerns 2. may lack important signals b. Types of cell cultures Primary cultures: directly from living organism ☺ reflect function of real organism Primary cells eventually die off; # some cell types poor dividers (nerve) time Cell lines: derived from primary cells (not clones) ☺ altered genes ⇒ immortality •isolated spontaneous mutation •tumor cells # •viral oncogene transfection Cell lines time 3.051J/20.340J 2 2. Assays of Cell Function Adhesion Migration Proliferation Differentiation (change in phenotype, ex. blood stem cells ⇒ leukocytes) Secretion (protein production) a. Cell Adhesion Assays Importance: cell adhesion is necessary for many other cell functions— provides biophysical and biochemical stimulation Sedimentation assay: 1. Cell type seeded onto surface of interest at given density (#/area) for a specified time in vitro 2. Surface is gently washed, and remaining cells counted, e.