Outline - Chapter 8 - Chapter 8 (pages 501-531) A stem cell...

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Chapter 8 (pages 501-531) A stem cell story ( http://www.eurostemcell.org/Outreach/Film/filmeng.html ) - 15 min movie 1. Isolating cells and grouping them in culture – why? Isolate cells to isolate specific proteins, or processes or to culture these cells to create many of them - Tissues –heterogeneous cell populations so, to make a homogenous population of cells, - Cells attached to each other and to the extracellular matrix have to be broken apart o In Animal tissues proteolytic enzymes (trypsin and collagenase) are used to disrupt the extracellular matrix and cell-cell junction chelating agents (EDTA) chelate or bind the Ca 2+ on which cell to cell adhesion depend on o plant tissues pectinase proteolytic enzyme for plants that degrades pectin - Separation of cells o Antibody (coupled to a fluorescent dye) binding to a specific cell types FACS: antibody with fluorescent dye specifically binds to the surface of only one cell type in the tissue, then, labeled cells can be sorted from unlabeled cells in FACS Cells pass through a laser beam as it monitors for fluorescence. Droplets with single cells are given either a positive charge (no fluorescence antibody present) or negative charge (fluorescence antibody present). Then droplets deflected by electric field into separate collection tubes. More efficient than microdissection o Microdissection Tissue gets coated with thin plastic film then a infrared laser is used to cut out the cells of interest then ejected into a container for further analysis Permits isolation of even a single cell from a tissue sample - Cell culture: tend to get “culture shock” and die because of replicative cell senescence and because check point mechanisms arrest the cell cycle (cells stop dividing after a finite number of divisions because of telomere shortening because of stopped production of telomerase) o Primary cultures – cells obtained directly from the tissues o Secondary cultures – recultured cells from primary culture o Transformed cell lines: normal cells that are transformed with a tumor- inducing virus or chemical
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Most easily generated from cancer cells that can inactivate checkpoint mechanisms and become “immortal” Grow without attaching to surface Proliferate more o Stem cells (ES Cells: embryotic stem cells): From inner cell mass of early embryo Proliferates indefinitely If these cultured cells put back into environment, can give rise to all cell types in body and adopt character and behavior of that cell Important not to transplant ES cells by themselves into adults because can cause tumors called teratomas Body will reject ES cells if they don’t match genetically o Cloning – reproductive and therapeutic: a clone is a set of individuals (cells) that are genetically identical because they have descended from
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This note was uploaded on 04/12/2008 for the course ENG 316K taught by Professor Kruppa during the Spring '08 term at University of Texas at Austin.

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Outline - Chapter 8 - Chapter 8 (pages 501-531) A stem cell...

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