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ctech chp 3 - Coggle Diagram

- - - - rich in proteins, act as buffer against changes
  - - - influence survival differentiation and proliferation
  - - - growth hormone-nitrogenic effect
        
        hydrocortisone-promote cell attachment and proliferation
  - - - detoxifying role , protect cells from damage by shear force
- - - - lab has problem maintaining a few cell lines
- - - - cell culture lacks original blood circulation
    - - strict aseptic working conditions
        
        frequent sub-culturing
        
        change media often
        
        mutated, specific cell functions lost or altered
  - - - divide for limited number of times, senesce after ~30 cycles of division
      - maintain some degree of differentiation
    - - propagated indefinitely through transformation(spontaneous like cancer or chemically/virally induced)
        
        retained little of the original in vivo characteristics
        
        limitless availability
  - - - disadvantage:can only be maintained in vitro for a limited period of time
        
        advantage: retain differential characteristics of the cell in vivo
      - cell suspension is reseeded/subcultured into new flask-passage
- - - - adaptation-cells lost extracellular matrix during trypsinization, reproduce it
      - attachment to substrata(a prerequisite for cell proliferation esp for anchorage dependent cells) so glycoproteins are produced
      - no cell division
      - cell spreading occurs
      - amount of dna polymerase increases-new dna and structural proteins synthesised
    - - less motility,isnt any empty space to spread and proliferate
        
        cells occupy less SA of substrate, presents less of its own surface to medium
        
        increase synthesis of specialised proteins(instead of structural proteins), growth rate reduced
        
        cell number in [steady state, cell growth=cell death
  - - - Inorganic salts: balanced salt solution (BSS)
        
        BSS is made of inorganic salts to form a solution
        
        Eagle’s MEM with Hank’s salts (HBSS)
        • or Eagle’s MEM with Earle’s salts (EBSS)
        
        functions
        
        maintain physiological pH
        
        – maintain osmotic pressure of the cells
        
        – maintain membrane potential ,
        
        – act as cofactors in enzyme reactions
        
        – aid in cell attachment (adhesion molecules need Mg and Ca
        
        application
        
        dilute concentrated amino acids or vitamins to form complete media
        
        3) For washing or dissection (cheaper than using complete medium
        
        4) For short incubation (30-60minutes
        
        energy sources
        
        glucose
        
        l-glutamine:an amino acid supplied for protein synthesis-provide major proportion of energy required by some cells
        
        • Defined as a combination of inorganic salts (such as Na,K,Mg,Ca,Cl,SO4,PO4,HCO3)
      - Buffer system: bicarbonate
      - Energy sources: carbohydrates
      - Proteins and peptides
      - Lipids (Fatty acids)
      - Amino Acids
        
        most animal cells require essential amino acids-those not synthesised by the body
        
        nonessential aas compensate for particular cell type's incapacity to make them, or they are lost by leakage in medium
      - Vitamins
        
        cell growth and multiplication
        
        precursor for metabolic cofactors
      - Accessory factors
      - Phenol red
- - - - should be aliquoted before refreezing
      - repeated freezing and thawing will lead to flocculent precipitation of proteins(fibrin threads)
        small amt can be used]
        
        prolonged incubation at 37c-small opaque dots(calcium phosphate) under the microscope-still can be used