Soldevila G, Buscema M, Marini V, Sutton R, James R F, Bloom S R, Robertson R P, Mirakian R, Pujol-Borrell R, Bottazzo G F
Department of Immunology, University College and Middlesex School of Medicine, London, UK.
J Autoimmun. 1991 Jun;4(3):381-96. doi: 10.1016/0896-8411(91)90154-5.
At present, only islet cell lines of animal origin have been successfully generated (e.g. RIN, HIT). A fully differentiated human beta cell line would be advantageous for diabetes research. We now report the generation of a human endocrine pancreatic cell line obtained by transfection using a plasmid containing the early region of SV40 viral DNA. Viral integration and transcription was assessed by Southern and Northern blotting. This cell line has been growing continuously for more than 2 years and maintains several of the characteristics of the parental cells from which they were generated. The presence of Neuron Specific Enolase, Protein Gene Product 9.5, cytokeratin, microvilli, cytoplasmic electrodense granules and the secretion of insulin, glucagon and somatostatin supports the neuroendocrine origin of this cell line. However, hormone production progressively decreased and finally stopped at passage 8. Flow cytometric analysis showed that HLA expression in this cell line is readily induced by IFN-gamma and modulated by TNF-alpha. The establishment of this human endocrine cell line indicates the feasibility of immortalizing human islets by transfection with viral oncogenes. To obtain a fully differentiated cell line it may be necessary to use other DNA constructs which immortalize the cells without fully transforming their phenotype.
目前，仅成功构建了动物源胰岛细胞系（如RIN、HIT）。一个完全分化的人β细胞系对糖尿病研究将是有利的。我们现在报告通过用含有SV40病毒DNA早期区域的质粒转染获得的人内分泌胰腺细胞系的构建。通过Southern和Northern印迹法评估病毒整合和转录。该细胞系已连续生长超过2年，并保留了其来源的亲代细胞的一些特征。神经元特异性烯醇化酶、蛋白基因产物9.5、细胞角蛋白、微绒毛、细胞质电子致密颗粒的存在以及胰岛素、胰高血糖素和生长抑素的分泌支持了该细胞系的神经内分泌起源。然而，激素产生在传代8时逐渐减少并最终停止。流式细胞术分析表明，该细胞系中的HLA表达可被IFN-γ轻易诱导，并受TNF-α调节。这个人内分泌细胞系的建立表明通过用病毒癌基因转染使人胰岛永生化是可行的。为了获得一个完全分化的细胞系，可能有必要使用其他DNA构建体，这些构建体在不完全改变其表型的情况下使细胞永生化。
