Bioreactor system

• US 5,081,035 A
• Filed: 02/11/1991
• Issued: 01/14/1992
• Est. Priority Date: 04/18/1988
• Status: Expired due to Fees

• Current Assignee: University of Michigan
• Original Assignee: University of Michigan
• Inventors: Midgley, A. Rees, Halberstadt, Craig R.
• Primary Examiner(s): Hill, Jr., Robert J.
• Assistant Examiner(s): BEISNER, WILLIAM H
• Application Number:
  US07/654,418
• Time in Patent Office: 337 Days
• Field of Search: 435/284-286, 435/288, 435/311, 435/313, 435/240.242, 210/321.73, 210/321.78, 210/321.77, 210/321.80, 210/500.23
• US Class Current: 435/297.4
• CPC Class Codes: C12M 23/24 Gas permeable parts
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 29/12 Pulsatile flow
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• CPC Class Codes: C12M 35/08 Chemical, biochemical or biological means, e.g. plasma jet, co-culture
• CPC Class Codes: C12M 41/00 Means for regulation, monitoring, measurement or control, e.g. flow regulation controlling or regulating chemical, physical or physicochemical processes B01J19/0006; heating or cooling apparatus for laboratory use B01L7/00; electro optical investigation of individual particles, flow cytometers G01N15/14; automatic analysis G01N35/00; controlling or regulating in general G06N
• CPC Class Codes: C12M 41/32 of substances in solution

Abstract

A system for continuous perfusion culturing of anchorage-dependent and anchorage-independent mammalian cell lines, inter alia, wherein in vivo capillary bed conditions are simulated by providing a bioreactor having a growth chamber wherein the cells are grown between a multitubular array comprising at least three functionally separate, inert non-degradable and structurally strong tubes or sets of tubes. A constant nutrient gradient is maintained along the entire length of the tubes by perfusing medium through the tubes at a flow rate which is sufficient to expose all areas of the chamber to fresh medium by convective forces rather than diffusion. Oxygen transfer and removal of toxic wastes is improved by the invention herein. In a preferred embodiment, porous inert expanded Teflon tubes are employed having an inner diameter of approximately between 1 and 2 millimeters.

Claims

1. An apparatus for maintaining and culturing cells in vitro therein, the apparatus comprising:

a housing;

a first permeable tube arranged in said housing for supplying a liquid medium, said first permeable tube being arranged in said housing along a first path therein which has a first predetermined length, said first permeable tube having first and second ports at respective ends thereof and positioned and arranged so as to communicate with the exterior of said housing, said first path being substantially randomly disposed within the housing;

a second permeable tube arranged in said housing for receiving the liquid medium supplied by said first permeable tube and containing a product produced by the cells, said second permeable tube being arranged in said housing along a second path having a second predetermined length, said second path being maintained throughout said second predetermined length of said second path in close proximity to substantially the entire first predetermined length of said first path by being intertwined therewith so as to provide a varying distance between said first and second paths throughout said first and second predetermined lengths, and having first and second ports at respective ends thereof and positioned and arranged so as to communicate with the exterior of said housing, whereby a convective flow of the liquid medium is maintained in a region in the vicinity of said first and second paths; and

enclosure means for providing a fluid-tight closure for said housing, said first permeable tube and said second permeable tube being disposed within said fluid-tight closure of said enclosure means.

2. The apparatus of claim 1 wherein there is further provided pulsed-pressure medium supply means for supplying said liquid medium to said first permeable tube.

3. The apparatus of claim 1 wherein at least one of said first and second permeable tubes has an internal diameter of approximately between 1 and 2 millimeters.

4. The apparatus of claim 1 further including spool means for supporting said first and second permeable tubes so as to be substantially randomly disposed within said housing.

5. The apparatus of claim 1 wherein there is further provided a third permeable tube for propagating a gas having a predetermined composition, wherein said third permeable tube is intertwined with said first permeable tube and said second permeable tube.

6. The apparatus of claim 5 wherein said first, second, and third permeable tubes are arranged to be intertwined in a braided manner with one another.

7. The apparatus of claim 5 wherein at least one of said first, second, and third permeable tubes is provided with a nonporous tubing affixed at an end thereof for connecting said one of said first, second, and third permeable tubes to a connector.

8. The apparatus of claim 5 wherein said third permeable tube is formed of a gas-permeable silicone rubber.

9. The apparatus of claim 5 wherein said first, second, and third permeable tubes have an intertubular distance which is less than approximately 2 millimeters.

10. The apparatus of claim 2 wherein said third permeable tube is formed of expanded polyfluorinated hydrocarbon.

11. The apparatus of claim 5 wherein at least one of said first, second, and third permeable tubes is formed of a microporous polyfluorinated hydrocarbon material.

12. The apparatus of claim 11 wherein said microporous polyfluorinated material has an overall porosity of approximately 70%.

13. The apparatus of claim 12 wherein said microporous polyfluorinated material has pores therein having a pore size of approximately between 2.0 and 4.0 microns.

14. An apparatus for maintaining and culturing cells in vitro therein, the apparatus comprising:

a housing;

a first permeable tube formed of a microporous polyfluorinated hydrocarbon material having an internal diameter of approximately between 1 and 2 millimeters for supplying a liquid medium;

a second permeable tube formed of a microporous polyfluorinated hydrocarbon material having an internal diameter of approximately between 1 and 2 millimeters for receiving a product fluid formed at least in part of the liquid medium supplied by said first permeable tube whereby a convective flow of the liquid medium is maintained in a region in the vicinity of said first and second permeable tubes;

a third permeable tube formed of a microporous polymeric material for conducting a gas-containing fluid having a predetermined composition, said first, second and third permeable tubes being intertwined with respect to one another so as to have a distance therebetween which varies along their respective lengths;

spool means for supporting said first, second, and third permeable tubes so as to be substantially randomly disposed within said housing;

enclosure means for providing a fluid-tight closure for said housing said first, second, and third permeable tubes, and said spool means; and

first, second, and third pairs of fluid coupling ports, each such pair of fluid coupling ports being coupled to a respective one of said first, second, and third permeable tubes at respective distal ends thereof, so as to provide fluid communication with the exterior of said enclosure means.