animal cell biotech

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54 Terms

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uses of cultured cells over intact organisms
isolate single cells to grow into clone colony
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can manipulate cells
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examine effect of treatments of intracellular events
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advantages of using cultured cells over intact organisms
more homogeneous than cells in tissues
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can control experimental conditions
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no ethical issues
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requirements for growing cells in culture
rich growth media
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serum with growth factors
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sterile conditions
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correct pH and osmolarity
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special solid surface
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how is primary culture generated
established from animal tissues - cells extracted from tissue using a protease
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how do we grow cells to form a primary cell line
cells grow in a monolayer in dish until space is filled
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small number of cells need to be passaged and transferred to another dish to continue cell line growth
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senescence in animal cells
most cells from an animal grow and divide for a limited period - around 50 doublings - then die
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cell line def
culture of cells with indefinite life span
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what happens when some cells escape senescence
said to be transformed - arises spontaneously through genetic changes
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these form a cell line
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characteristics of transformed cell lines
immortal
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anchorage independent
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less need for growth factors
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motile
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no contact inhibition of growth
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tumourigenic
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less differentiated
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2 uses of animal cells
protein production
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tissue engineering
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polyclonal antibody
from several different B cell clones that recognise different epitopes
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monoclonal antibody
from a single B cell clone that recognises a single epitope
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how can two cells be used to make a hybrid
mouse tumour cells in culture are fused with mouse spleen cells to form hybridomas
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these are used to produce monoclonal antibodies
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uses of monoclonal antibodies
research - measuring/detecting specific proteins/antigens
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disease diagnosis
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therapy
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stages in tissue engineering
design scaffold from biodegradable substance
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expand cells from biopsy
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seed scaffold with cells and allow grow
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implant into patient
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pros of ESCs
pluripotent so can form any cell in body
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cons of ESCs
not genetically identical to patient
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difficult to culture
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ethical issues
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adult SCs pros
genetically identical to patient
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no ethical issues
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adult SCs cons
form limited number of cell types
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not available for all tissues
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how are iPS cells made
skin cells removed from patient
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reprogram cells so they become induced pluripotent stem cells
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treat them so they differentiate into specific cell type & return to patient
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pros/cons of iPS cells
fewer ethical issues & can use patients own cells
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technically challenging & risk of cancer
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how does direct reprogramming work
reprogramming of one somatic cell type directly into another cell type
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doesn't need IPS state
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achieved by expression of key TFs - uses viruses and CRISPR-cas9