High Density Culture Using Macroporous Microcarrier

Shirokaze, J.; Nogawa, Makoto; Ogura, Ryuma

doi:10.1016/b978-0-7506-1845-8.50063-6

Cited by 4 publications

(3 citation statements)

References 5 publications

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“…Cytopore is one of the excellent porous microcarriers made of cellulose for animal cell culture, which possesses almost all characteristics of an ideal cell culture support, as described by Griffiths (Griffiths, 1990). It can be used for both anchorage dependent and suspension cells, is capable of long-term continuous operation with low-serum/serum-free media, has good mechanical stability, and considerable volumetric scale-up potential (Shirokaze, 1994). In this study, we report that a rCHO cell line which expresses prourokinase was cultivated on Cytopore porous microcarriers in a 30l Biostat UC stirred tank reactor with a spinfilter to retain cells.…”

Section: Introductionmentioning

confidence: 99%

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Hu¹,

Xiao²,

Huang³

et al. 2000

Cytotechnology

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A recombinant DNA CHO cell line secretingurokinase-type plasminogen activator (u-PA) wascultivated with Cytopore cellulose porousmicrocarriers in a 30l Biostat UC stirred tankreactor. After 26 days of culture, using a spinfilter toretain cells in bioreactor, the cell density couldreach 1.33 x 10(7) ml(-1). The maximal u-PAactivity in supernatant was 7335 IU.ml(-1), and204l supernatant containing 7.1 g u-PA was harvested.After 100 days of culture with 0.1% fetal bovineserum medium, a modified cell retention system whichcan be washed-out backward, substituted thespinfilter to prevent filter clogging. The maximalcell density was over 10(7) ml(-1), the maximalu-PA activity in supernatant reached 6250IU.ml(-1), and 1604l supernatant containing about51 g u-PA was harvested. Compared to perfusionculture, batch medium-replaced culture could raiseutilizing efficiency of the medium, increase cell specificproductivity and improve the quality of the product which wasnot steady in a 37 degrees C environment. Cells can movefrom seed porous microcarriers occupied by cells tovacant microcarriers spontaneously, withouttrypsinization, and continue to grow until all microcarriers contained cells. It shows that Cytoporeporous microcarriers are very useful and convenient toscale up cultivation step by step.

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Section: Introductionmentioning

confidence: 99%

Untitled

Hu¹,

Xiao²,

Huang³

et al. 2000

Cytotechnology

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“…The development of microcarrier culture by van Wezel in 1967(Van Wezel, 1967 rendered the large-scale industrial culture of anchorage dependent cells possible for the production of vaccine and interferon. In 1980s, a great advance of this procedure was achieved by the development of a series of porous microcarriers (Lobby, 1990;Gotoh, 1993;Shirokaze, 1994). In a previous paper we reported that using porous microcarrier Cytopore, we successfully cultivated a genetically-engineered CHO cell line and some hybridomas to produce pro-UK and McAB (Xiao, 1996).…”

Section: Introductionmentioning

confidence: 99%

Untitled

Xiao¹,

Huang²,

Li³

et al. 1999

Cytotechnology

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Using porous microcarrier Cytopore and a low-serum medium supplement BIGBEF-3, we have successfully cultivated recombinant CHO cell line CL-11G producing prourokinase and hybridomas producing anti-prourokinase monoclonal antibody in Celligen 1.5 or 5 L bioreactor. The cell density obtained ranged from 1 to 2 x 107 cells mL-1. The yields of prourokinase and monoclonal antibody increased with increasing cell density. As the cells could spontaneously release from and reattach to porous microcarriers, it was very easy to scale-up the cultivation. Thus the bead to bead cell transfer method has been used to scale up the cultivation of CL-11G cells to a 20 L reactor-scale for the pilot production of prourokinase, and also to scale-up the culture of hybridomas for the production of monoclonal antibody for the purification of prourokinase.

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“…Another system is the Biott spinner (Able K K, Japan), which is equipped with a coil of a porous Teflon tube for aeration. This system allows the cell numbers to be counted during the cultivation and it is suitable for suspension as well as microcarrier cultures (Shirokaze et al, 1993). This system is equipped with pH and oxygen electrodes.…”

Section: Introductionmentioning

confidence: 99%

The Super-Spinner: A low cost animal cell culture bioreactor for the CO2 incubator

et al. 1994

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The production of small quantities of monoclonal antibodies and recombinant proteins was carried out using a new low cost production system, the Super Spinner. Into a 1 1 standard Duran | flask a membrane stirrer equipped with a polypropylene hollow fiber membrane was installed to improve the oxygen supply by bubble-free aeration. The aeration was facilitated by using the CO 2 conditioned incubator gas, which was pumped through the membrane stirrer via a small membrane pump. The maximal oxygen transfer rate (OTRmax) of the Super Spinner was detected. For this purpose one spinner flask was equipped with an oxygen electrode. The OTRma ~ was measured by the dynamic method. The ratio of membrane length to culture volume was adapted corresponding to the oxygen uptake rate of the cells according to the desired cell density. A balanced nutrient supply resulted in an optimal formation and yield of products.

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High Density Culture Using Macroporous Microcarrier

Cited by 4 publications

References 5 publications

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The Super-Spinner: A low cost animal cell culture bioreactor for the CO2 incubator

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