Method and apparatus for growing biomass particles

• US 5,654,197 A
• Filed: 08/06/1993
• Issued: 08/05/1997
• Est. Priority Date: 01/17/1992
• Status: Expired due to Term

• Current Assignee: Merck Serono SA (Merck & Co., Inc.)
• Original Assignee: Applied Research Systems ARS Holding NV (Merck & Co., Inc.)
• Inventors: Jem, Kwan-Min, Cevey, Pierre-Francois
• Primary Examiner(s): BEISNER, WILLIAM H
• Application Number:
  US08/102,098
• Time in Patent Office: 1,460 Days
• Field of Search: 435/240.1, 435/240.22, 435/240.23, 435/240.24, 435/240.25, 435/240.46, 435/243, 435/261, 435/284-286, 435/311, 435/313, 435/315, 435/316, 435/81 3, 210/800, 210/513, 210/616, 210/538, 210/540, 210/150, 210/151, 210/615, 422/101, 422/225, 422/228, 422/269, 422/271
• US Class Current: 435/383
• CPC Class Codes: C12M 27/02 Stirrer or mobile mixing elements
• CPC Class Codes: C12M 33/22 Settling tanks; Sedimentation by gravity settling tanks per se B01D21/02

Abstract

The present invention provides an improved method and apparatus for growing biomass particles, particularly microcarrier-bound cells, in an agitated suspension culture vessel in which fresh culture medium is added and spent culture medium is withdrawn continuously or semi-continuously. The improvement comprises withdrawing the spent culture medium through a particle settling chamber located within the vessel and at least partially immersed in the agitated culture medium therewithin. The particle settling chamber comprises a hollow container with a bottom opening through which biomass particles, such as microcarrier-bound cells, settle by gravity back into the agitated culture medium and a top opening through which particle-free spent culture medium is withdrawn form the vessel. The settling chamber is configured such that the fluid velocity of culture medium entering the settling chamber through the bottom opening is significantly less than the biomass particle settling velocity.

Claims

1. A method for growing biomass particles in an agitated suspension culture vessel, comprising concurrently

(A) adding fresh culture medium to an agitated suspension culture vessel containing biomass particles;

and

(B) withdrawing spent culture medium from said agitated suspension culture vessel containing biomass particles through a side perforated particle settling chamber which is at least partially immersed in an agitated culture medium within said vessel containing biomass particles, wherein

(i) said particle settling chamber comprises a hollow container with a bottom opening through which said biomass particles settle by gravity back into the agitated culture medium and a top opening through which the spent culture medium is withdrawn from the vessel;

(ii) said chamber is configured such that a fluid velocity of the culture medium entering said container through said bottom opening is significantly less than a settling velocity of the biomass particles; and

(iii) said hollow container has a plurality of holes disposed along the sides thereof, said holes being sized and located so as to allow culture medium and biomass particles to enter said container while avoiding significant agitation therewithin and minimizing withdrawal of biomass particles through the top opening thereof.

2. The method of claim 1, wherein said hollow container is

(a) closed to the atmosphere within said vessel;

(b) said bottom opening being significantly larger in cross-sectional area than said top opening; and

(c) said top opening communicates exclusively outside said vessel.

3. The method of claim 2, wherein said hollow container has an essentially cylindrical shape.

4. The method of claim 2, wherein said hollow container or an uppermost portion of said hollow container has a cone or inverted funnel shape, wherein the narrow portion of said cone or inverted funnel shape defines said top opening.

5. The method of claim 1, wherein said top opening is open to the atmosphere within said vessel and the spent culture medium is withdrawn from said vessel through a dip tube inserted through said top opening partially into said hollow container so as to contact the culture medium therewithin.

6. The method of claim 5, wherein said hollow container has one wall which is formed by a portion of the wall of said culture vessel and another wall which is formed by a wall piece that extends inwardly from said culture vessel wall portion and abuts the culture vessel wall portion along two separate and approximately vertical lines.

7. The method of claim 1, wherein said biomass particles comprise microcarrier-bound cells.

8. An apparatus for growing biomass particles, comprising an agitated suspension culture vessel, a particle settling chamber disposed within said vessel such that it will be at least partially immersed in agitated culture medium during use, wherein

(i) said particle settling chamber comprises a hollow container with (1) a bottom opening through which said biomass particles settle by gravity back into an agitated culture medium and (2) a top opening through which spent culture medium is withdrawn from the vessel;

(ii) said chamber is configured such that a fluid velocity of the culture medium entering said container through said bottom opening is significantly less than a settling velocity of the biomass particles; and

(iii) said hollow container has a plurality of holes along the sides thereof, said holes being sized and located so as to allow the culture medium and biomass particles to enter said container while avoiding significant agitation therewithin and minimizing withdrawal of biomass particles through the top opening thereof.

9. The apparatus of claim 8, wherein said top opening communicates solely outside said vessel and said bottom opening is significantly larger than said top opening, wherein during use said hollow container is closed to the atmosphere within said vessel.

10. The apparatus of claim 9, wherein said hollow container has an essentially cylindrical shape.

11. The apparatus of claim 9, wherein said hollow container or an uppermost portion of said hollow container has a cone or inverted funnel shape, wherein the narrow portion of said cone or inverted funnel shape defines said top opening.

12. The apparatus of claim 8, further comprising a dip tube which extends from a point outside said vessel through said top opening partially into said hollow container, such that during use said dip tube contacts the culture medium within said hollow container, wherein said top opening is larger in cross-sectional area than said dip tube.

13. The apparatus of claim 12, wherein said hollow container has one wall which is formed by a portion of the wall of said culture vessel and another wall which is formed by a wall piece that extends inwardly from said culture vessel wall portion and abuts the culture vessel wall portion along two separate and approximately vertical lines.

14. An apparatus according to claim 8, wherein said hollow container comprises a grid located at or near the bottom opening, said grid reducing or preventing agitation within said hollow container.

15. An apparatus according to claim 14, wherein said grid is configured so as to readily allow biomass particles to pass through during use.