Nongenetic Techniques for Isolating Fusion Products between Different Cell Types
Until recently, virtually all investigations into the behavior of cell hybrids have employed genetic selection techniques, such as the hypoxanthine-aminopterin-thymidine (HAT) system (Littlefield, 1964), to isolate the cell fusion products. In this approach, one starts with two cell lines, one of which lacks enzyme A and the other of which lacks enzyme B. After fusing the two cell types together, they are placed in a specially designed culture medium in which only cells that have both enzymes are able to grow. Under these conditions, neither parental cell line is able to divide. However, since the cell line that lacks enzyme A has enzyme B and vice versa, fusion products between the two do contain both enzymes and the cell hybrids are able to multiply. Although systems based on this approach have been extremely useful, there are a variety of practical and theoretical problems associated with their use. Obviously, only special cell lines that lack the necessary enzymes can be used. Although in some cases this problem can be surmounted by mutagenizing and isolating cells carrying the required defect, this can only be done with immortal cell lines. The cultured life span of normal diploid cells (Hayflick, 1965) is limited, so that by the time mutant clones are isolated they are essentially senescent.
KeywordsCell Hybrid Fusion Product Diphtheria Toxin Selective System Isotonic Medium
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