Murata M, Togami M, Tawara Y, Onodera K
Tsukuba Research Laboratory, Japan Synthetic Rubber Co., Ltd., Ibaraki.
Cytotechnology. 1993;13(2):143-8. doi: 10.1007/BF00749941.
We recently developed a new dialysis culture system (termed LIFROC-device) for the cultivation of lymphokine-activated killer cells (Murata et al., 1990, 1991). In the present study, we applied the LIFROC-device (400 ml culture vessel) to the cultivation of mammalian cells for the production of biologically active substances. We cultured mouse-mouse hybridoma TP-709, secreting anti-tisse plasminogen activator (tPA) monoclonal antibody (mAb), recombinant CHO GT19, secreting hGH, and human melanoma Bowes cells, secreting tPA. With the LIFROC-device, TP-709 grew to a maximal cell density of 3.8 x 10(6) cells/ml and and produced 480 micrograms/ml (192 mg in total) of mAb. GT19 reached a cell density of 2.2 x 10(6) cells/ml and produced 302 micrograms/ml (120 mg in total) of hGH. Bowes cells expanded to 4.4 x 10(6) cells/ml and secreted 8.5 micrograms/ml (3.3 mg in total) of tPA. The protein concentration in the culture broths of the LIFROC-device became 7-200 times higher than that of batch culture. Thus, the LIFROC-device can be applied to protein production as well as cell growth with high efficiency.
我们最近开发了一种用于培养淋巴因子激活的杀伤细胞的新型透析培养系统（称为LIFROC装置）（村田等人，1990年，1991年）。在本研究中，我们将LIFROC装置（400毫升培养容器）应用于哺乳动物细胞的培养，以生产生物活性物质。我们培养了分泌抗组织型纤溶酶原激活剂（tPA）单克隆抗体（mAb）的小鼠-小鼠杂交瘤TP-709、分泌hGH的重组CHO GT19以及分泌tPA的人黑色素瘤Bowes细胞。使用LIFROC装置，TP-709生长至最大细胞密度为3.8×10⁶个细胞/毫升，并产生了480微克/毫升（总计192毫克）的mAb。GT19达到细胞密度为2.2×10⁶个细胞/毫升，并产生了302微克/毫升（总计120毫克）的hGH。Bowes细胞扩增至4.4×10⁶个细胞/毫升，并分泌了8.5微克/毫升（总计3.3毫克）的tPA。LIFROC装置培养液中的蛋白质浓度比分批培养高7至200倍。因此，LIFROC装置可高效应用于蛋白质生产以及细胞生长。
