Cell Bio Notes

Cell Bio Notes - Chapter 8: Manipulating Proteins, DNA,...

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Section 1: Isolating cells and growing them in culture To obtain as much info as possible about cells in a tissue, biologists have developed ways f dissociating cells from tissues and separating them according to type. Cells can be isolated from intact tissues - disrupt the extracellular matrix and cell-cell junctions that hold cells together - most general cell sepration techniquye uses an antibody coupled to a fluorescent dye to label specific cells. An antibody is chosen that specifically binds to the surface of only one cell type in the tissue - they can then be seprated from the unlabaled ones in an electronic fluorescence active cell sorter. Cells can be grown in culture - cells grown in culture provide a more homogenous population of cells from which to extract material Eukaryotic cell lines are a widely used source of homoegenous cells - cell cultures obtained by disrupting tissues usually tend to die because the cell has turned off telomerase which is why telemores shorten and stop dividing. - To immortalize these cells, one has to do more than add telomerase, one must inactivetae the checkpoint mechanisms. This is done by introducing cancer-promoting oncogenes. - Transformed cell lines: cells regenerated from cancer cells Embryonic stem cells could revolutionize medicine - Using ES to create a potentially inexhaustible supply of cells that might be used to replace and repair damaged mature human tissue( ie skeletal muscles that degenerate in MD victims) - Major problem: transplated cells differ genetically from the cells of the patient Somatic cell nuclear transplationg may provide a way to generate personalized stem cells - Reproductive cloning: cloning of entire multicellular orgs. - The need for 2 parents and sexual union is bypassed by somatic cell nuclear transplantation. - Procedure begins with an unfertilized egg cell. The nucleus of this haploid cell is sucked out and replaced by a nuclear from a regular diploid somatic cell. the hybrid cell is develops and is put into the uterus of the foster mother. It will be genetically identical to the adultat that donated the diploid cell. Hybridoma cell lines are factories that produce monoclonal antibodies - Antibodies are produced by inoculating animals with the protein of interest and isolating the antiobodies specific to that protein from the serum of the animal.
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Section 2: Purifying proteins Purifying a single type of protein from 1000s is a formidable challenge. Recombinant DNA technology can simplify this task by “tricking” cells into producing large quantities of a given protein, thereby making its purification much easier. Cells can be separated into their component fractions
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This note was uploaded on 03/08/2011 for the course BIO 110 taught by Professor Rexach during the Spring '08 term at UCSC.

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Cell Bio Notes - Chapter 8: Manipulating Proteins, DNA,...

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