BIOL 3450 Lecture Notes - Lecture 4: Cell Culture, Senescence, Glutamine
14 January 2019
BIOL 3450
Chapter 4 – Culturing and Visualizing Cells
Cell Culture Media
o Salts and buffering agents
o Nutrients
▪ Glucose
▪ Vitamins – organic compounds body needs but can’t make itself
▪ Essential amino acids – bodies can’t synthesize all amino acids; we need to take
in via other sources
▪ Glutamine – cells can make but if given extra, it helps; more readily accessible
source of Nitrogen
o Growth factors
▪ Serum from fetal calves
▪ Serum free media has insulin, transferring + growth factors
Difference between Transformed or Stem Cell Lines
o Primary Cultures – cells directly from body
▪ Require attachment matrix
▪ Senescence after limited number of cell divisions
▪ Contact inhibited; grow as monolayers on culture dishes → stop growing when
they come in contact with other cells
▪ Differentiated cell types may require 3D structure support or “feeder” layers of
cells
o Transformed (Cancer) or Stem Cell Lines
▪ Can grow in liquid suspension
▪ “immortal” – can be cultured indefinitely
▪ Not contact inhibited; form 3D mounds or balls → just keep dividing
▪ Relatively undifferentiated
Light Microscopy
o Resolution (D)
▪ Ability to distinguish between two closely positioned objects
• D = 0.61 theta / N sin a
• N – refractive index of medium between specimen and objective lens
• Theta – wavelength of light
• Alpha – half angle of cone of light that enters objective lens
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