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lecture13 - 1 3.051J/20.340J Lecture 13 Quantifying Cell...

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1 3.051 J / 20 .340 J 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 / eventually die off; # / some cell types poor dividers (nerve) Primary cells time Cell lines: derived from primary cells (not clones) altered genes immortality isolated spontaneous mutation tumor cells viral oncogene transfection # Cell lines time
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2 3.051 J / 20 .340 J 2. Assays of Cell Function ¾ Adhesion ¾ Migration ¾ Proliferation ¾ Differentiation (change in phenotype, ex. blood stem cells leukocytes) ¾ Secretion (protein production) a. Cell Adhesion Assays Importance: cell adhesion is necessary for many other cell functions— provides biophysical and biochemical stimulation ¾ 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.g., by optical microscopy or Coulter counter (cells detached, suspended & “count” by electrical resistance change thru narrow channel) % seeded adhered treated surface TCPS (control) 3.For testing cell resistance, serum-containing medium is a stricter test than cells seeded in protein-free solution. Note: plasma vs. serum plasma—liquid portion of blood (cells removed) serum—plasma with coagulants (e.g., FGN) removed
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3 3.051 J / 20 .340 J Sedimentation assays give no info . on strength of adhesion.
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