Culturing method, system and apparatus for cell culture
First Claim
1. A culturing method of living cells comprising the steps of:
- withdrawing a cell culture broth from a culturing apparatus;
bringing the withdrawn cell culture broth into contact with a membrane for precision filtration wherein said membrane separates the cells present in the culture broth from the filtrate;
while filtering the cell culture broth through the use of said membrane, bringing a gas which is non-toxic to the cells present in the culture broth into contact with a surface of the membrane immersed in said culture broth to thereby prevent the adherence and propagation of said cells to the membrane surface;
rupturing the foam occurring on the liquid surface as a result of the addition of the gas to the culture broth by a defoaming layer comprised of a hydrophobic material disposed on the liquid surface;
returning the cells thus separated from the filtration to said culturing apparatus; and
,withdrawing the separated filtrate from said culturing apparatus.
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Accused Products
Abstract
A culturing method and system are disclosed in which a cell culture broth in withdrawn from a culturing apparatus to a filteration apparatus. The withdrawn broth in brought into contact with a membrane for precision filteration. Then a non-toxic gas is brown into the broth, while filtering the broth to separate the cells and breaking foams occurring on the surface of the broth due to gas bubbling. The separated cells are returned to the vessel and the filterate is withdrawn from the filteration apparatus.
The method and system have such advantages that a perfusion culture of a long period of time can be realized because the clogging of the filteration membrane is effectively prevented.
19 Citations
10 Claims
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1. A culturing method of living cells comprising the steps of:
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withdrawing a cell culture broth from a culturing apparatus; bringing the withdrawn cell culture broth into contact with a membrane for precision filtration wherein said membrane separates the cells present in the culture broth from the filtrate; while filtering the cell culture broth through the use of said membrane, bringing a gas which is non-toxic to the cells present in the culture broth into contact with a surface of the membrane immersed in said culture broth to thereby prevent the adherence and propagation of said cells to the membrane surface; rupturing the foam occurring on the liquid surface as a result of the addition of the gas to the culture broth by a defoaming layer comprised of a hydrophobic material disposed on the liquid surface; returning the cells thus separated from the filtration to said culturing apparatus; and
,withdrawing the separated filtrate from said culturing apparatus. - View Dependent Claims (2, 3, 4, 5)
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6. A culturing system for living cells, which comprises:
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a) a culturing apparatus having a vessel for culturing cells in the suspension state in a culture broth; b) means for supplying culturing cells into the vessel; c) means for supplying a medium into the vessel thereby forming the culture broth containing the culturing cells; d) means for withdrawing a part of the culture broth from the vessel; e) means for transferring the withdrawn culture broth to a filtration apparatus having a membrane for precision filtration wherein the membrane separates the cells in the culture broth from the filtrate; f) means for bringing a gas which is non-toxic to the cells present in the culture broth into contact with the surface of the membrane of the filtration apparatus to thereby prevent adherence and propagation of the cells to the membrane; g) means for rupturing the foam occurring on the surface of the culture broth in the filtration apparatus due to the addition of the gas to the culture broth; h) means for returning the cells separated from the filtrate in the filtration apparatus to the vessel; and
,i) means for withdrawing the filtrate from the filtration apparatus. - View Dependent Claims (7, 8, 9, 10)
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Specification