| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1985: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1984: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
We tried to get somatic cell hybrids which were able to produce homogeneous(monoclonal) hormone and grow well in vitro. Cell fusions were performed by polyethylene glycol. After selection of hybrids, cultured medium, hybrids, and extract of the hybrids were examined for hormone production by radioimmunoassay(RIA), enzyme-linked immunosorbent assay(ELISA), and immunofluorescence(FA).
RIA showed that none of hybrids between murine fibroblastic cells( <LmTK^-> ) and the insulinomas or rat pituitary tumor cells(GH3:growth hormone(GH) producer) produced insulin nor GH. Hybrids between the insulinoma cells and GH3 did not produce insulin nor GH. Meanwhile, hybrids between same hormone producing cells(e.g. insulinoma x insulinoma, GH3 x GH3) were found to produce either inslin or GH.
Next, cell fusions were performed between <LmTK^-> and rat primary pancreatic islets,and 275 hybrids were obtained. FA revealed that some of them had somatostatin-like substance or glucagon-like substance 9 months after the cell fusion. Then, we focused our interest on the somatostatin-like substance to quantify it. This substance, however, was undetectable by RIA. To characterize the difference in results between FA and RIA for somatostatin, anti-somatostatin antibody used for FA was analyzed by the 2 absorption tests using 2 different antigens. These experiments supported the production of somatostatin-like substance by the hybrid. It might be possible that anti-somatostatin antibody which was used for RIA is not able to recognize the antigenic determinant of the somatostatin-like substance in the hybrid. Futher investigations are required to make the issue clear.
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