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CELL PRODUCT OF MAMMALIAN INSULIN-PRODUCING CELLS AND METHODS FOR USING THE SAME

机译:哺乳动物胰岛素产生细胞的细胞产物及其使用方法

摘要

The technical result of the invention is to simplify the technology of obtaining insulin-producing cells, obtaining at least 70% of functionally active insulin-producing cells in cell culture, that underwent differentiation. The method comprises obtaining epithelial progenitor cells and their subsequent differentiation into pancreatic cells, capable to glucose-sensitive insulin secretion in which pancreatic differentiation is performed in two stages: (a) at the first stage cells are differentiated within 4-15 days in a culture medium containing at least serum of a mammal, glutamine, epidermal growth factor, transferrin, sodium selenite, retinoic acid, isoproterenol; (b) at the second stage, cells are differentiated within 4-15 days in a culture medium containing at least serum of a mammal, glutamine, epidermal growth factor, retinoic acid, nicotinamide, hepatocyte growth factor, dexamethasone; moreover, the cultivation in both stages is carried out in gas atmosphere of 5% CO2 at 37°C. Group of inventions includes cell product of insulin-producing cells of a mammal, and a method of differentiation of pancreatic epithelial progenitor cells of mammals, including humans, as well as a method for replacement therapy of diabetes mellitus using cell product.
机译:本发明的技术结果是简化获得胰岛素产生细胞的技术,即获得在细胞培养物中经历分化的至少70%的具有功能活性的胰岛素产生细胞。该方法包括获得能够对葡萄糖敏感的胰岛素分泌的上皮祖细胞及其随后的分化为胰腺细胞,其中胰腺分化分为两个阶段:(a)在第一阶段,在培养物中4-15天内分化细胞至少含有哺乳动物血清,谷氨酰胺,表皮生长因子,转铁蛋白,亚硒酸钠,视黄酸,异丙肾上腺素的培养基; (b)在第二阶段,在至少包含哺乳动物血清,谷氨酰胺,表皮生长因子,视黄酸,烟酰胺,肝细胞生长因子,地塞米松的培养基中的4-15天内分化细胞;此外,两个阶段的培养均在37℃的5%CO 2的气体气氛中进行。本发明的组包括哺乳动物的胰岛素产生细胞的细胞产物,以及包括人在内的哺乳动物的胰腺上皮祖细胞分化的方法,以及使用该细胞产物替代糖尿病的方法。

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