BIOL 303 Lecture Notes - Lecture 2: Differential Centrifugation, Buoyancy, Column Chromatography
Chapter 8
• Recall the origins, most common types, and purpose of culturing primary and
immortalized cell lines (chapter 8, slides 4-6).
• Primary cultures = cells grown directly from a tissue.
• Brain tissue is chopped up and plated on a dish.
• Can only survive for weeks.
• This is due to the shortening of telomeres as replication continues.
• Retinal ganglion cells, coming from the brain.
• Immortalized cell lines = primary cells that divide indefinitely.
• Example: through addition of telomerase in somatic cells or a cancerous
mutation
• 3T3: Fibroblast (mouse), HeLa: Epithelial cell (human), 293:
kidney(human); transformed with adenovirus, CHO: Ovary (Chinese
hamster). All of these are common cell lines used.
• Uses a virus strategy or cancerous mutation.
• Organoids = Coaxing cells to form a pseudo-organ.
• Recognize the differences between and understand the reasons for using FACS, velocity
sedimentation, equilibrium sedimentation, column chromatography, and
immunoprecipitation (chapter 8, slides 7-12).
• FACS: separates cells based on their fluorescence automatically via a detector.
• Velocity sedimentation: Based on size and shape.
• Equilibrium sedimentation: Based on buoyant density. Uses a sucrose gradient.
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