Distributed perfusion bioreactor system for continuous culture of biological cells

Patent metadata

Field	Value
Publication number	US-10214718-B2
Application number	US-201414900946-A
Country	US
Kind code	B2
Filing date	Jul 1, 2014
Priority date	Jul 1, 2013
Publication date	Feb 26, 2019
Grant date	Feb 26, 2019

Abstract

Embodiments provide a distributed bioreactor system in which a plurality of modular bioreactors are operated in parallel to produce and maintain a biological cell culture. A central or nurse perfusion bioreactor produces and maintains a cell culture and transfers portions of the cell culture to a plurality of modular peripheral perfusion bioreactors, each of which produces and maintains the cell culture in turn. In order to prevent contamination and facilitate segregation of particular peripheral bioreactors, the distributed system is configured such that one-way fluid communication is established from the central/nurse bioreactor to each of the peripheral bioreactors while maintaining fluid isolation among the peripheral bioreactors. Each modular peripheral bioreactor unit has a plug-and-play configuration and may be plugged into or otherwise coupled to the central/nurse bioreactor to scale up the overall size of the cell culture without requiring sterilization or redesign or reconfiguration of the system.

First claim

What is claimed is:

1. A distributed bioreactor system for producing and maintaining a continuous biological cell culture, the system comprising:
a nurse perfused bioreactor configured to produce and maintain a cell culture, the nurse perfused bioreactor coupled to a nurse perfusion device;
a plurality of peripheral bioreactors configured to produce and maintain a cell culture in parallel;
an individual and distinct peripheral perfusion device positioned proximal to and coupled to each respective individual and distinct peripheral bioreactor of the plurality of peripheral bioreactors;
one or more control devices coupled to and configured to control the nurse perfusion device and the peripheral perfusion devices;
a multi-way manifold coupled to the nurse perfused bioreactor; and
a plurality of one-way fluid conduits configured to provide fluid communication from the nurse perfused bioreactor to the plurality of peripheral bioreactors, each one-way fluid conduit coupling the nurse perfused bioreactor to one of the peripheral bioreactors using the multi-way manifold;
wherein configuration of the multi-way manifold and the one-way fluid conduits enables transfer of the cell culture from the nurse bioreactor to at least two of the peripheral bioreactors and maintains fluid isolation among each of the peripheral bioreactors.

2. The distributed bioreactor system of claim 1 , wherein each peripheral bioreactor and its corresponding one-way fluid conduit are pre-assembled to form a sterile modular unit suitable for coupling to the nurse bioreactor via the multi-way manifold to scale up a volume of the cell culture of the distributed bioreactor system; optionally
wherein each peripheral bioreactor has the same working volume;
wherein the plurality of peripheral bioreactors comprises 3, 4 or 5 peripheral bioreactors;
wherein the one or more control devices are configured to independently control one or more operational characteristics of the bioreactors, wherein the operational characteristics are selected from the group comprising a rate of agitation, a temperature, a pH, an oxygen level, a viable cell density based on a cell concentration measurement, a rate of perfusion, a weight, a volume, a metabolic flux, a metabolic rate, and combinations thereof; and/or
wherein the multi-way manifold is switchable among the plurality of one-way fluid conduits, the system further comprising a manifold control device configured to activate the multi-way manifold to sequentially feed cells in the cell culture from the nurse bioreactor to at least two of the peripheral bioreactors; and/or wherein the manifold control device configures the multi-way manifold to automatically close off fluid communication between the multi-way manifold and a first peripheral bioreactor upon detecting a process deviation or a contamination in the first peripheral bioreactor.

3. The distributed bioreactor system of claim 1 , further comprising:
a plurality of cell concentration probes coupled to and configured to measure capacitance of the cell cultures of the nurse and peripheral bioreactors; and
a plurality of volume sensing probes coupled to and configured to measure a volume, a liquid level or a liquid weight of the cell cultures of the nurse and peripheral bioreactors;
wherein the one or more control devices are configured to control in real-time an operational state of a first bioreactor among the nurse and peripheral bioreactors based at least on a first cell concentration based on the first cell concentration measured by a first capacitance probe or other cell concentration sensors coupled to the first bioreactor and/or based on a first volume, a first liquid level or a first liquid weight measured by a first volume sensing probe coupled to the first bioreactor; optionally
wherein the one or more control devices are configured to:
upon inoculation of the cell culture in the first bioreactor, shut off a medium inlet port and a medium outlet port of the first bioreactor to maintain the cell culture in the first bioreactor at a substantially constant volume during a batch state;
upon determining that a cell concentration of the cell culture in the first bioreactor has reached a first predefined cell concentration threshold, open the medium inlet port to introduce a cell growth medium into the first bioreactor to maintain the cell concentration substantially at the first cell concentration threshold during a fed-batch state;
upon determining that a volume of the cell culture in the first bioreactor has reached a first predefined volume threshold, open the medium inlet port and the medium outlet port to control the first perfusion device to retain cells within the bioreactor, introduce the cell growth medium into the first bioreactor and release spent cell growth medium from the first bioreactor, while maintaining the volume of the cell culture substantially at the first volume threshold during a perfused batch state;
upon determining that a cell concentration of the cell culture in the first bioreactor has reached a second predefined cell concentration threshold, open the medium inlet port and control the first perfusion device to retain cells within the bioreactor, introduce the cell growth medium into the first bioreactor and release spent cell growth medium from the first bioreactor to maintain the cell concentration substantially at the second cell concentration threshold during a perfused fed-batch state; and
upon determining that a volume of the cell culture in the first bioreactor has reached a second predefined volume threshold, open the medium inlet port and the medium outlet port and control the first perfusion device to retain cells within the bioreactor, introduce the cell growth medium into the first bioreactor and release spent medium from the first bioreactor to maintain a volume of the cell culture substantially at the second volume threshold while maintaining the cell concentration constant or not using a bleeding of cells.

4. The distributed bioreactor system of claim 1 , further comprising:
a plurality of cell concentration probes coupled to and configured to measure cell concentration of the cell cultures of the nurse and peripheral bioreactors;
wherein the one or more control devices are configured to automatically control in real-time an operational state of a first bioreactor among the nurse and peripheral bioreactors based at least on a first cell concentration based on the first cell concentration measured by a first cell concentration probe coupled to the first bioreactor; optionally
wherein the one or more control devices are configured to automatically change a first operational state of the first bioreactor to a second operational state when the first cell concentration reaches a first predefined cell concentration threshold.

5. The distributed bioreactor system of claim 4 , wherein the first operational state is a batch state in which the cell concentration of the cell culture increases while the volume is maintained at a substantially constant level; and
wherein the second operational state is a fed-batch state in which the volume of the biological cell culture is adjusted to maintain the cell concentration at a substantially constant level.

6. The distributed bioreactor system of claim 4 , wherein the first operational state is a perfused batch state in which a first perfusion device is operated to retain cells within the bioreactor, introduce a cell growth medium into the first bioreactor and remove spent medium from the first bioreactor, while maintaining the volume of the cell culture at a substantially constant level; and
wherein the second operational state is a perfused fed-batch state in which the first perfusion device is operated to retain cells within the bioreactor, introduce

Assignees

• Berteau Olivier
• Yoon Seongkyu
• Sergeant David
• Poskin Thierry
• Univ Massachusetts
• Apicells Inc

Inventors

• Berteau Olivier
• Yoon Seongkyu
• Sergeant David
• Poskin Thierry

Classifications

• C12M41/44

  of volume or liquid level · CPC title

• C12M41/48 Primary

  Automatic or computerized control (automatic analysis G01N35/00) · CPC title

• C12M29/10

  Perfusion · CPC title

• C12M23/28

  disposable or single use · CPC title

• C12M23/40

  Manifolds; Distribution pieces (fluid transfer means B01L3/563) · CPC title