Hollow fiber cell culture device and method of operation

• US 4,804,628 A
• Filed: 08/19/1987
• Issued: 02/14/1989
• Est. Priority Date: 10/09/1984
• Status: Expired due to Term

• Current Assignee: Biovest International, Inc.
• Original Assignee: Endotronics Incorporated
• Inventors: Walker, Robert D., Gruenberg, Micheal L., Cracauer, Ray F.
• Primary Examiner(s): Tarcza, John
• Application Number:
  US07/088,463
• Time in Patent Office: 545 Days
• Field of Search: 435/240.242, 435/310, 435/284, 435/285, 435/948, 210/321.1, 210/323.2, 210/335, 210/433.2, 210/650, 210/651, 210/806, 436/178
• US Class Current: 435/400
• CPC Class Codes: C12M 25/10 Hollow fibers or tubes hollow fiber modules in general B01D63/02
• CPC Class Codes: C12M 29/16 Hollow fibers hollow fiber modules in general B01D63/02
• CPC Class Codes: C12M 29/18 External loop; Means for reintroduction of fermented biomass or liquid percolate loop type reactors for chemical or physical processes B01J19/2435
• 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 41/40 of pressure

Abstract

An improved cell culturing device includes a hollow fiber cartridge having a shell and a plurality of capillaries extending through the shell with at least some of the capillaries having selectively permeable walls. A cell culturing space is located between the shell and capillaries. The improvement includes a chamber containing a second medium supply fluidly connected to the cell culturing space. A pressurizing system pressurizes the medium within the chamber to a level higher than the level of medium flowing through the lumen of the capillaries. A valving mechanism alternatively and selectively restricts flow of medium between the chamber and the cell culturing space through first and second conduits such that circulation is effected in the cell culturing space.

Claims

1. An improved cell culture device for in vitro cell growth, the device including a shell having first and second ends, first and second ports, and a plurality of capillaries extending between the first and second ends within the shell with at least some of said capillaries having selectively permeable walls, and cell culturing space being defined between the shell and the capillaries with the first and second ports in fluid communication therewith, a first supply of medium being delivered into the capillaries at a first selected pressure, the improvement comprising:

a medium expansion chamber fluidly connected to the first and second ports and containing a second supply of medium at a second pressure;

valving means for selectively restricting flow of the second supply of medium through the first and second ports into the culturing space, wherein the valving means operates such that flow of the second supply of medium is altered through the first and second ports to effect medium circulation within cell culturing space and enhance diffusion of waste products out of the culturing space and diffusion of medium components into the culturing space while retaining cells and desired cell products;

pressurizing means for keeping the second pressure in the chamber approximately constant and maintaining the second pressure independently of the first pressure; and

means for selectively raising and lowering the first pressure in the capillaries with respect to the second pressure in the chamber effecting circulation within the cell culturing space.

2. The device of claim 1 wherein the chamber is fluidly connected to the first and second ports of the shell by first and second conduits and wherein the valving means includes first and second valves controlling flow through the first and second conduits, respectively.

3. The device of claim 1 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 50,000 Daltons.

4. The device of claim 3 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 15,000 Daltons.

5. A cell culture device for in vitro cell growth, the device comprising:

a hollow fiber cartridge having a shell with a plurality of capillaries extending between a fluid input end and a fluid output end and defining an extracapillary space between the shell and the capillaries with at least some of said capillaries having selectively permeable walls and the shell having first and second spaced-apart ports communicating with the extracapillary space;

delivery means for transporting to the capillaries a first supply of medium and including first means for maintaining the first supply of medium in the capillaries at a first pressure that is selectively raised and lowered;

a chamber fluidly connected to the first and second ports containing a second supply of medium and including second means for maintaining the second supply of medium at a selected second pressure;

valving means for selectively controlling flow of the second supply of medium from the chamber through the first and second ports; and

wherein the delivery means alternatively raises and lowers the first pressure in the capillaries above and below the second pressure in the chamber and wherein the first means for maintaining and the second means for maintaining control the first and second pressures independently of each other and in cooperation with the valving means such that circulation of medium within the extracapillary space is effected.

6. The device of claim 5 wherein the chamber is fluidly connected to the first and second ports of the shell by first and second conduits and wherein the valving means includes first and second valves restricting flow in the first and second conduits, respectively.

7. The device of claim 5 wherein the second means for maintaining includes a mechanism for delivering gas under pressure into the chamber.

8. The device of claim 5 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 50,000 Daltons.

9. The device of claim 8 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 15,000 Daltons.

10. A method for culturing cells in a hollow fiber cartridge having a shell and a plurality of capillaries extending through the shell, and defining a cell culturing space between the capillaries and shell with the cell culturing spce being fluidly connected to an expansion chamber through a first and second port, each with a valve, the capillaries being provided with a first supply of medium by a delivery system the method comprising:

maintainng a first pressure through the first supply of medium in the capillaries;

maintaining a second pressure through a second supply of medium in the cell culturing space independently of the first pressure; and

alternatively increasing and decreasing the first pressure within the capillaries above and below the second pressure within the cell culturing space so that flow of medium from the cappilaries into the cell culturing space is effected when the first pressure in the capillaries is greater than the cell culturing space and flow of medium is effected from the cell culturing space in the capillaries when the first pressure is less than the second pressure, the alternating of the first pressure above and below the second pressure producing circulation in the cell culturing space.

11. The method of claim 10 wherein the first pressure is increased and decreased sufficiently to cause ultrfiltration of components from the medium through the capillary walls into the cell culturing space and ultrafiltration of components deleterious to cells in the cell culturing space into the lumen of the capillaries, respectively.

12. The method of claim 11 and further including establishing an upper level sensing point and a lower level sensing point in the expansion chamber and sensing when the medium has reached the upper level and the lower level;

and increasing the pressure within the capillaries when the low level has been sensed and decreasing the pressure in the capillaries when the high level has been sensed.

13. An improved cell culture device for in vitro cell growth, the device including a shell having first and second ends, first and second ports, and a plurality of capillaries extending between the first and second ends within the shell with at least some of said capillaries having selectively permeable walls, and a cell culturing space being defined between the shell and the capillaries with the first and second ports in fluid communiation therewith, a first supply of medium being delivered into the capillaries at a first selected pressure, the improvement comprising:

a chamber fluidly connected to the first and second ports and containing a second supply of medium independent of the first supply of medium for delivery at a second selected pressure;

valving means for selectively restricting flow of the second supply of medium through the first and second ports into the culturing space, wherein the valving means is operated such that flow of the second supply of medium is alternated through the first and second ports to effect circulation within the cell culturing space and enhance diffusion of waste products out of the culturing space and diffusion of medium components into the culturing space;

second pressurizing means for keeping the second pressure approximately constant;

upper and lower level sensors disposed at a selected upper and lower level within the chamber to sense a lower level and an upper level of medium; and

wherein the delivery means is selectively controlled along with the valving means to raise the first pressure in the capillaries with respect to the second pressure in the chamber when a low level is sensed and to lower the first pressure in the capillaries with respect to the second pressure in the chamber when a high level is sensed effecting circulation within the cell culturing space.

14. The device of claim 13 wherein the chamber is fluidly connected to the first and second ports of the shell by first and second conduits and wherein the valving means includes first and second valves controlling flow through the first and second conduits, respectively.

15. The device of claim 13 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 50,000 Daltons.

16. The device of claim 15 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 15,000 Daltons.

17. A cell culture device for in vitro cell growth, the device comprising:

a hollow fiber cartridge having a shell with a plurality of capillaries extending between a fluid input end and a fluid output end and defining an extracapillary space between the shell and the capillaries with at least some of said capillaries having selectively permeable walls and the shell having first and second spaced-apart ports communicating with the extracapillary space;

delivery means for transporting to the capillaries and for selectively raising and lowering a first pressure of a first supply of medium in the capillaries;

a chamber fluidly connected to the first and second ports containing a second supply of medium;

.

upper and lower level sensors disposed to sense an upper and lower level of the medium within the chamber, respectively;

valving means for selectively controlling flow of the second supply of medium from the chamber through the first and second ports;

means for pressurizing the chamber at a second pressure and maintaining the second pressure independently of the first pressure; and

wherein the delivery means alternatively raises the first pressure in the capillaries above the second pressure in the chamber when a low level of medium is sensed in the chamber and lowers the first pressure in the capillaries below the second pressure in the chamber when a high level of medium is sensed in the chamber, said raising and lowering of first pressure in the chamber being done in cooperation with the valving means so that circulation of medium within the extracapillary space is effected.

18. The device of claim 17 wherein the chamber is fluidly connected to the first and second ports of the shell by first and second conduits and wherein the valving means includes first and second valves restricting flow in the first and second conduits, respectively.

19. The device of claim 17 wherein the means for pressurizing the chamber includes a mechanism for delivering gas under pressure into the chamber.

20. The device of claim 17 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 50,000 Daltons.

21. The device of claim 20 wherein the permeable walls of the capillaries have a selective molecular weight cut-off of less than approximately 15,000 Daltons.