Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, 194, Machida-shi, Tokyo, Japan.
Cytotechnology. 1991 Jan;5(Suppl 2):17-34. doi: 10.1007/BF00573878.
We investigated the basic technology of cell culture conditions for production of useful substances such as cytokines, and related proteins produced by Namalwa cells. Namalwa cells (Klein, 1972), human B lymphoblastoid cells, were used for large scale production of alpha-interferon (Klein, 1979). Namalwa KJM-1, a subline of Namalwa cells, adapted to serum- and albumin-free medium, can grow at a high density above 1 × 10(7) cells/ml in suspension mode by the use of a perfusion culture system, Biofermenter(™), containing a cone-type cell-sedimentation column as cell separator (Sato, 1983).Several kinds of cytokine cDNA can be introduced and expressed in Namalwa KJM-1 cells (Miyaji, 1990a,b,c). Some of these were produced in large quantities by use of a gene amplification method with dhfr (Miyaji, 1990c), even though the Namalwa KJM-1 cells contained endogenous dhfr genes. For stable production of the target protein, Namalwa KJM-1 cells are very useful host cells, because they have no effective endogenous protease activity in the conditioned medium.Using Biofermenter with micro-silicone fibers and a dialysis system, the specific productivity of the target proteins was not depressed at a high cell density.
我们研究了细胞培养条件的基本技术，以生产有用物质，如细胞因子和 Namalwa 细胞产生的相关蛋白质。Namalwa 细胞（Klein，1972）是人类 B 淋巴母细胞，用于大规模生产α干扰素（Klein，1979）。Namalwa KJM-1 是 Namalwa 细胞的一个亚系，适应无血清和无白蛋白培养基，可以在悬浮模式下通过使用灌注培养系统（Biofermenter(™)）在高密度下生长，该系统含有一个锥形细胞沉降柱作为细胞分离器（Sato，1983）。Namalwa KJM-1 细胞可以引入和表达几种细胞因子 cDNA（Miyaji，1990a，b，c）。其中一些通过使用 dhfr 的基因扩增方法大量生产（Miyaji，1990c），即使 Namalwa KJM-1 细胞含有内源性 dhfr 基因。为了稳定生产目标蛋白，Namalwa KJM-1 细胞是非常有用的宿主细胞，因为它们在条件培养基中没有有效的内源性蛋白酶活性。使用带有微硅纤维的 Biofermenter 和透析系统，在高细胞密度下，目标蛋白的特异性生产率不会受到抑制。
