Phytohemagglutinin-enhanced hybrid colony formation by cells from aged mice: Expression of th 4mod-1 mouse locus in human-mouse hybrids |
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Authors: | Patricia G West Wilber W Baker |
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Institution: | (1) Department of Biology, Villanova University, 19085 Villanova, Pennsylvanna |
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Abstract: | Summary Cells from a continuous human line and freshly isolated cells from old adult mice heterozygous at theMod-1 locus were fused in the presence of polyethylene glycol (PEG). The production of hybrid cells, as a function of PEG concentration
in the presence and absence of phytohemagglutining (PHA), was measured by cell survival and proliferation on selective medium.
The incorporation of PHA into the fusion mixture allowed cell fusion to take place at nontoxic concentrations of PEG. PHA
increased the frequency of cell fusion and increased the production of viable hybrid cells from 138- to over 2800-fold depending
on cell type. The results suggest that the procedure may have broad application in promoting the fusion of cells sensitive
to PEG. Clones were analyzed for isozymes of malic enzyme and glucose-6-phosphate dehydrogenase. The expression of the gene
encoding X-linked mouse glucose-6-phosphate dehydrogenase confirmed that the cells were hybrids. These cells lost other mouse
isozymes rapidly. In those clones in which the mouse malic enzyme gene was expressed, the product ofMod-1
α was detected significantly more frequently than that ofMod-1
b. |
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Keywords: | cell fusion polyethylene glycol phytohemagglutinin allozyme malic enzyme aging |
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