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Engineering research on the production of useful biologicals by mass culture of animal and plant cells

动植物细胞大量培养生产有用生物制品的工程研究

基本信息

批准号：
02453113
负责人：
FUNATSU Kazumori
金额：
$ 3.07万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

项目摘要

1. We could succeed in regulating a dissolved oxygen concentration (DO) of the medium in PID manner by using DO electrode, DO converter, PID controller and mass flow controller, in which the gas flow rate was used as the manipulated variable. By using the DO control system, it was found that there were optimum DO level in cell growth (3.0ppm) and beta-carotene production (0.05ppm) of carrot cells in an air-lift column with draft tube. Highest productivity (0.24mug/10^8cells/day) was obtained in DO-stat (0.15ppm) semi-continuous culture of carrot cells and this value was 7.5 time higher than that in the usual batch culture. We also succeeded in high density culture (1.1X10^8cells/cm^3) of animal cells by regulating a out-put DO level (more than 5.8ppm) of polyurethane foam (PUF) packed-bed culture system.2. The three-dimensional laminar flow of culture medium in an agitated vessel with 2-bladed paddle imoeller was simulated numerically by finite element method. The Carreau model was used as the constitutive equation. We succeeded in obtaining the fields of the velocity and the shear stress under a certain agitation rate (10-60 rpm).3. Adult rat hepatocytes formed spherical multicellular aggregates (spheroids, about 100mum in diameter) when they were cultured in the pores of PUF. Highly differentiated hepatic functions were maintained in the spheroid, These spheroids were partly attached and immobilized in the pores of PUF. We designed a PUF/spheroid packed-bed culture system. Albumin and urea production activities of hepatocyte/spheroid immobilized in the PUF packed-bed were higher than those of usual hepatocyte/monolayer culture in petri dish, and these activities were maintained for 26 days. These results indicated that the PUF/spheroid packed-bed culture system provided a promising prototype of hybrid type artificial liver support system.

1.通过采用DO电极、DO转换器、PID控制器和质量流量控制器，以气体流量为被控变量，实现了对培养液溶解氧浓度（DO）的PID调节。通过对胡萝卜细胞生长和β-胡萝卜素生产的DO控制系统的研究，发现在导流管气升塔中，胡萝卜细胞生长和β-胡萝卜素生产的最佳DO水平分别为3.0ppm和0.05ppm。在DO-stat（0.15ppm）半连续培养中，胡萝卜细胞的生产率最高（0.24 μ g/10^8个细胞/天），是常规分批培养的7.5倍。通过调节聚氨酯泡沫（PUF）填充床培养系统的输出DO水平（大于5.8ppm），成功地实现了动物细胞的高密度培养（1.1 × 10^8个/cm ^3）.采用有限元法对双叶桨式搅拌槽内培养基的三维层流流动进行了数值模拟。Carreau模型被用作本构方程。在一定的搅拌速率（10-60 rpm）下，成功地获得了速度场和剪切应力场.当成年大鼠肝细胞在PUF孔中培养时，它们形成球形多细胞聚集体（球状体，直径约100 μ m）。高分化的肝脏功能维持在球状体中，球状体部分附着并固定在PUF的孔隙中。我们设计了一个PUF/球形填充床培养系统。固定在PUF填充床中的肝细胞/球体的白蛋白和尿素产生活性高于通常的肝细胞/单层培养皿中的那些，并且这些活性维持26天。结果表明，PUF/球形填充床培养系统为复合型人工肝支持系统提供了一个有希望的原型。