Continuous culture apparatus with mobile vessel, allowing selection of fitter cell variants and producing a culture in a continuous manner

Abstract

A method and device for growing plant, animal or stem cells in a continuous manner.

Description

FIELD OF THE INVENTION [0001] The described invention provides a method and a device that allow selection of living cells, with increased rates of reproduction and specific metabolic properties, in a liquid or semi-solid medium. For the process of selection (adaptive evolution), genetically variant organisms (mutants) arise in a population and compete with other variants of the same origin. Those with the fastest rate of reproduction increase in relative proportion over time, leading to a population (and individual organisms) with increased reproductive rate. This process can improve the performance of organisms used in industrial processes or academic purpose. The present invention utilizes a continuous culture apparatus to achieve the viable production of living cells, for example, yeast, plant cells, animal cells or stem cells. The present invention may be used to produce an active ingredient or biologic that is produced by the living cells. The active ingredient or biologic may ...

AI Technical Summary

Benefits of technology

[0015] The present invention is designed to achieve these goals without any fluid transfer, including sterilization or rinsing functions. This represents a specific advantage of the present invention with respect to prior art in so far as it avoids the hazards and difficulties associated with sterilization and rinsing, including containment and complex fluid transfers involving caustic solvents.

[0016] Continuous culture is achieved inside a flexible sterile tube filled with growth medium. The medium and the chamber surface are static with respect to each other, and both are regularly and simultaneously replaced by peristaltic movement of the tubing through “gates”, or points at which the tube is sterilely subdivided by clamps that prevent the cultured cells from moving between regions of the tube. UV gates can also (optionally) be added upstream and downstream of the culture vessel for additional security.

[0017] The present method and device are also an improvement over prior art insofar as they continually, rather than periodically, select against adherence of dilution-resistant variants to the chemostat surfaces, as replacement of the affected surfaces occurs in tandem with the process of dilution.

[0018] The tube is subdivided in a transient way such that there are regions that contain saturated (fully grown) culture, regions that contain fresh medium, and a region between these two, wherein one or more chambers referred to as growth or culture chambers are present to form a growth chamber region in which grown culture is mixed with fresh medium to achieve dilution. The gates are periodically released from one point on the tube and replaced at another point, such that grown culture along with its associated growth chamber surface and attached static cells, is removed by isolation from the growth chamber and replaced by both fresh medium and fresh chamber surface.

Problems solved by technology

This process is usually done manually, with considerable labor investment, and is subject to contamination through exposure to the outside environment.

Serial culture is also inefficient, as described in the following paragraph.

Traditional chemostats are unable to sustain long periods of selection for increased reproduction rate, due to the unintended selection of dilution-resistant (static) variants.

This method and device achieves the goal, but requires independent complex manipulations of several fluids within a sterile (sealed) environment, including one (NaOH) which is both very caustic and potentially very reactive, quickly damaging valves, and posing containment and waste-disposal problems.

The chemostatic device is also limited in that no provisions are made to provide a support for cells to grow on

There are some types of cells that are difficult to culture in large amounts due to the conditions the cells require to survive and grow.

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