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Research on mass culture of cell from the viewpoint of chemical engineering

从化学工程的角度进行细胞大量培养的研究

基本信息

批准号：
62550705
负责人：
FUNATSU Kazumori
金额：
$ 1.28万
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 (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62550705/
关键词：
High density mass cell culture Animal cell Polyurethane foam Packed bed culture Fibrinolytic enzyme Plant cell beta-carotene Air-lift column equipped with draft tube ポリウレタンフォームドラフトチューム付きエアーリフト槽カロチノイド色素大量培養 Vero細胞 MPC-11細胞マイ

项目摘要

1. High density culture of anchorage-dependent mammalian cell using polyurethane foam as a substratum for cell attachment and production of useful biologicals Anchorage-dependent mammalian cells were cultivated at high cell density in a novel culture system using polyurethane foam (PUF) as a substratum for cell attachment. PUF has a macroporous structure giving a high surface area to volume ratio. Monkey kidney cells (Vero) attached to the internal surface of PUF and grew to a high cell density (1.04x10^8 cells/cm^3 PUF) in PUF stationary cultures. Furthermore, Vero cells were found to produce urokinase stored in the cell. In PUF stationary culture, Vero cells produced the urokinase from the early logarithmic phase of cell growth. The specific activity of urokinase became highest in the late-logarithmic phase and attained to 70 I.U./mg-protein. This maximal specific activity was about three times as large as that in petri dish culture. These results suggested that the urokinase product … More ivity of Vero cells increased by high density cell culture using PUF.2. Liquid spouted bed culture of microencapsulated mammalian cells Microencapsulated MPC11 cells, which are anchorage-independent, were tried to propagate in a newly designed liquid spouted bed culture system. As the culture system was suitable for separation of medium and microcapsules, the medium could be circulated sufficiently to give nutrients, oxygen, etc. to microencapsulated cells and wash out waste. As a result, the maximum cell density in the microcapsules reached comparatively high 3.3x10^7 cells/ml.3. Production and purification of carotenoid pigments from high density culture of carrot cells A method for purification of carotenoid pigments from cultured carrot cells was established. The relation between cell growth and carotenoid production of carrot cells in shake culture and air-lift culture was studied by using the purification method. The results indicated that carotenoids were growth-dependently produced from the early logarithmic phase of cell growth, and that productivity decreased in the stationary phase, even if medium was exchanged in the sationary phase. Finally, 29.4 mu g/liter-medium of beta-carotene was obtained from high density cultured carrot cells using an air-lift column of 1 liter volume equipped with a draft tube. Less

1.以聚氨酯泡沫塑料为基质高密度培养哺乳动物细胞贴壁和生产有用生物活性物质在以泡沫塑料(PUF)为基质的新型细胞培养体系中，以高密度培养哺乳动物细胞。PUF具有大孔结构，具有较高的比表面积与体积比。猴肾细胞(Vero)附着在PUF的内表面，在PUF静止培养中生长到高密度(1.04x108个/cm3PUF)。此外，发现Vero细胞能产生储存在细胞内的尿激酶。在PUF静止培养中，Vero细胞在细胞生长的早期对数阶段产生尿激酶。尿激酶的比活力在对数末期达到最高，达到70IU./mg蛋白。这一最大比活力约为培养皿培养的3倍。这些结果表明，尿激酶产物…结论：1.用PUF进行高密度细胞培养可提高Vero细胞活性。微囊化哺乳动物细胞的液体喷动床培养微囊化MPC11细胞不依赖于锚定，尝试在一种新设计的液体喷动床培养系统中增殖。由于该培养体系适合于基质和微胶囊的分离，因此可以使基质充分循环，为微胶囊细胞提供营养、氧气等，并清洗废物。结果表明，微胶囊中的最大细胞密度达到了较高的3.3×10~(-7)个/毫升。胡萝卜细胞高密度培养中类胡萝卜素色素的制备与纯化建立了一种从胡萝卜细胞中分离纯化类胡萝卜素色素的方法。用纯化方法研究了摇瓶培养和气升培养条件下胡萝卜细胞生长与类胡萝卜素合成的关系。结果表明，类胡萝卜素的产生依赖于细胞生长的早期对数阶段，而在稳定期，即使在稳定期更换培养基，类胡萝卜素的产量也会下降。最后，采用1L气升柱和导流管，从高密度培养的胡萝卜细胞中获得29.4微克/升的β-胡萝卜素。较少