Cell culture - Cell culture Hybridoma production Research...

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Cell culture Hybridoma production
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Research Problem You are interested in researching pain regulation at the molecular level specifically by studying the action of the DREAM protein in mice. This protein is expressed in cells of the lumbar spinal cords. Your first task is to obtain primary cultures.
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Approach What potential technical difficulties you may encounter in obtaining a “pure” culture? What techniques can you use to obtain such cultures? What are the differences between primary and secondary cultures, and cell lines?
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Types of animal cells used in research: Primary cultures Obtained from tissues of freshly dissected animals Advantages: normal cells Disadvantages: finite number of cell divisions, sometimes hard to obtain a “pure”/homogenous cell culture. Why is this a problem? Secondary cultures Primary cultures are allowed to divide Advantages: cell differentiation can be studied. Ex: fibroblasts Disadvantages: finite number of cell divisions, cell isolation process can potentially eliminate factors (cell-cell interactions) needed for cell differentiation.
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