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Details for Patent: 6,391,638

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Details for Patent: 6,391,638

Title: Cell/tissue culturing device and method
Abstract:A disposable device and method for axenically culturing and harvesting cells and/or tissue in consecutive cycles. The device consists of a sterilisable disposable container which may be partially filled with a suitable sterile biological cell and/or tissue culture medium. The container has means for removing excess air and/or waste gases therefrom, and means for introducing inoculant and/or culture medium and/or additives therein. A reusable harvesting means enables harvesting of at least a portion of the medium containing cells and/or tissue when desired, thereby enabling the device to be used continuously for at least one subsequent consecutive culturing/harvesting cycle. The portion of medium containing cells and/or tissue remaining from a previously harvested cycle may serve as inoculant for a next culture and harvest cycle.
Inventor(s): Shaaltiel; Yoseph (Beit Hillel, IL)
Assignee: Metabogal, Ltd. (Kiryat Shemona, IL)
Filing Date:Feb 08, 1999
Application Number:09/246,600
Claims:1. A disposable device for axenically culturing and harvesting at least one of tissue and a quantity of cells in at least one cycle, said device comprising a sterilisable disposable non-rigid container which is at least one of transparent and translucent, said container having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissues and quantity of cells, when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining in said device from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided.

2. The device as claimed in claim 1, further comprising air inlet means for introducing sterile air in the form of bubbles into said culture medium through a first inlet opening, wherein said air inlet means is connectable to a suitable air supply.

3. The device as claimed in claim 1, said harvesting means comprising contamination prevention means for substantially preventing introduction of contaminants into said container via said harvesting means.

4. The device as claimed in claim 1, wherein said container is made from a non-rigid plastics material.

5. The device as claimed in claim 4, wherein said material is selected from the group comprising polyethylene, polycarbonate, a copolymer of polyethylene and nylon, PVC and EVA.

6. The device as claimed in claim 4, wherein said container is made from a laminate of more than one layer of said materials.

7. The device as claimed in claim 4, wherein said container is formed by fusion bonding two suitable sheets of said material along predetermined seams.

8. The device as claimed in claim 2, wherein said air inlet means comprises an air inlet pipe extending from said inlet opening to a location inside said container at or near the said bottom end thereof.

9. The device as claimed in claim 2, wherein said at least one air inlet means comprises a least one air inlet pipe connectable to a suitable air supply and in communication with a plurality of secondary inlet pipes, each said secondary inlet pipe extending to a location inside said container, via a suitable inlet opening therein, for introducing sterile air in the form of bubbles into said culture medium.

10. The device as claimed in claim 9, wherein said device comprises a substantially box-like geometrical configuration, having an overall length, height and width.

11. The device as claimed in claim 10, wherein the height-to-length ratio is between about 1 and about 3, and preferably about 1.85.

12. The device as claimed in claim 10, wherein the height to width ratio is between about 5 and about 30, and preferably about 13.

13. A disposable device for axenically culturing and harvesting at least one of tissue and a quantity of cells in at least one cycle, said device comprising a sterilisable disposable container which is at least one of transparent and translucent, said container having a top end and a bottom end, said device further comprising a substantially boxlike geometrical configuration, having an overall length, height and width, wherein the height-to-length ratio is between about 1 and about 3, said device further comprising a support aperture substantially spanning the depth of said device, said aperture adapted to enable said device to be supported on a suitable pole support wherein said container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) at least one air inlet means for introducing sterile air in the form of bubbles into said culture medium through a first inlet opening, said at least one air inlet means comprising at least one air inlet pipe connectable to a suitable air supply and in communication with a plurality of secondary inlet pipes, each secondary inlet pipe extending to a location inside said container, via a suitable inlet opening therein, for introducing sterile air in the form of bubbles into said culture medium, and

(iv) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissues and quantity of cells, when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided.

14. A disposable device for axenically culturing and harvesting at least one of tissue and a quantity of cells in at least one cycle, said device comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) at least one air inlet means for introducing sterile air in the form of bubbles into said culture medium through a first inlet opening, said at least one air inlet means comprising at least one air inlet pipe connectable to a suitable air supply and in communication with a plurality of secondary inlet pipes, each secondary inlet pipe extending to a location inside said container, via a suitable inlet opening therein, for introducing sterile air in the form of bubbles into said culture medium, said device further comprising a support structure for supporting said device; and

(iv) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissues and quantity of cells, when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided.

15. The device as claimed in claim 14, wherein said support structure comprises a pair of opposed frames, each said frame comprising upper and lower support members spaced by a plurality of substantially parallel vertical support members suitably joined to the said upper and lower support members.

16. The device as claimed in claim 15, wherein said plurality of vertical support members consists of at least one said vertical support member at each longitudinal extremity of the said upper and lower support members.

17. The device as claimed in claim 15, wherein said frames are spaced from each other by a plurality of spacing bars releasably or integrally joined to said frames.

18. The device as claimed in claim 16, wherein said spacing bars are strategically located such that the said device may be inserted and removed relatively easily from the said support structure.

19. The device as claimed in claim 15, wherein the said lower support member of each said frame comprises at least one lower support adapted for receiving and supporting a corresponding portion of the said bottom end of the said device.

20. The device as claimed in claim 19, wherein each said lower support is in the form of suitably shaped tab projecting from each of the lower support members in the direction of the opposed frame.

21. The device as claimed in claim 15, wherein said frames each comprise at least one interpartitioning means projecting from each frame in the direction of the opposed frame, for to pushing against the sidewall of said device at a predetermined position, such that opposed pairs of said interpartitioning means effectively reduce the width of the said device at said predetermined position.

22. The device as claimed in claims 21, wherein said interpartitioning means comprise suitable substantially vertical members spaced from the said upper and lower support members in a direction towards the opposed frame by means of suitable upper and lower struts.

23. The device as claimed in claim 14, wherein, said support structure may comprise a plurality of castors for transporting the said devices.

24. The device as claimed in claim 1, wherein said container comprises a suitable filter mounted on said gas outlet means for substantially preventing introduction of contaminants into said container via said gas outlet means.

25. The device as claimed in claim 1, wherein said container further comprises a suitable filter mounted on said additive inlet means for substantially preventing introduction of contaminants into said container via said additive inlet means.

26. A disposable device for axenically culturing and harvesting at least one of tissue and a quantity of cells in at least one cycle, said device comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissues and quantity of cells, when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle, said reusable harvesting means further comprising a contamination prevention means comprising a U-shaped fluid trap, wherein one arm thereof is aseptically mounted to an external outlet of said harvesting means by suitable aseptic connection means,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided.

27. A disposable device for axenically culturing and harvesting at least one of tissue and a quantity of cells in at least one cycle, said device comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) reusable harvesting means located at the bottom of said bottom end of said container, said harvesting means further comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissues and quantity of cells when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided.

28. A disposable device for axenically culturing and harvesting at least one of tissue and a quantity of cells in at least one cycle, said device comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) reusable harvesting means located near the bottom of said bottom end of said container, said harvesting means further comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said cells and tissues when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle,

wherein said harvesting means is positioned such that at the end of each harvesting cycle a remainder of said medium containing at least one of said cells and tissue automatically remains at said bottom end of said container up to a level below the level of said harvesting means, said remainder of said medium containing at least one of said cells and tissue, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided.

29. The device as claimed in claim 1, wherein said bottom end is substantially convex.

30. The device as claimed in claim 1, wherein said bottom end is substantially frusta-conical.

31. The device as claimed in claim 1, wherein said container comprises an internal fillable volume of between about 5 liters and about 200 liters.

32. A disposable device for axenically culturing and harvesting at least one of tissue and a quantity of cells tissue in at least one cycle, said device comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissue and quantity of cells when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided and said device further comprises suitable attachment means for attaching said device to a suitable support structure.

33. The device as claimed in claim 32, wherein said attachment means comprises a loop of suitable material preferably integrally attached to said top end of said container.

34. A battery of disposable devices, comprising at least two of said disposable devices for axenically culturing and harvesting at least one of tissue and a quantity of cells in at least one cycle, each of said devices comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissue and quantity of cells when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle, and

(iv) an air inlet means for introducing sterile air in the form of bubbles into said culture medium through a first air inlet opening, said air inlet means connectable to a suitable air supply,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided.

35. The battery as claimed in claim 34, wherein said devices are supported by a suitable support structure via the said attachment means of each said device.

36. The battery as claimed in claim 34, wherein the said gas outlet means of each said device is suitably connected to a common gas outlet piping which optionally comprises suitable means for preventing contaminants from flowing into said devices.

37. The battery as claimed in claim 36, wherein said means for preventing contaminants from flowing into said devices comprises a suitable filter.

38. The battery as claimed in claim 34, wherein the said additive inlet means of each said device is suitably connected to a common additive inlet piping having a free end optionally comprising suitable aseptic connecting means thereat.

39. The battery as claimed in claim 38, wherein said free end is connectable to a suitable supply of medium and/or additives.

40. The battery as claimed in claim 34, wherein the said harvesting means of each said device is suitably connected to a common harvesting piping having a free end optionally comprising suitable aseptic connecting means thereat.

41. The battery as claimed in claim 40, further comprising contamination prevention means for substantially preventing introduction of contaminants into said container via said common harvesting piping.

42. The battery as claimed in claim 41, wherein said contamination prevention means comprises a U-shaped fluid trap, wherein one arm thereof is free having an opening and wherein the other end thereof is aseptically mountable to said free end of said common harvesting piping via suitable aseptic connection means.

43. The battery as claimed in claims 42, wherein the said free end of said U-tube is connectable to a suitable receiving tank.

44. The battery as claimed in claim 34, wherein the said air inlet means of each said device is suitably connected to a common air inlet piping having a free end optionally comprising suitable aseptic connecting means thereat.

45. The battery as claimed in claim 44, wherein said free end is connectable to a suitable air supply.

46. A method for axenically culturing and harvesting at least one of tissue and a quantity of cells in a disposable device comprising the steps of:

a) providing said device which comprises a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and other sterile required additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

(iii) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a portion of said medium containing at least one of said tissue and quantity of cells when desired, thereby enabling said device to be used continuously for at least one further consecutive cycle, wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previously harvested cycle may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided;

b) providing axenic inoculant via said harvesting means;

c) providing sterile at least one of said culture medium and said additives via said additive inlet means;

d) optionally illuminating said container with external light means;

e) allowing at least one of said cells and tissue to grow in said medium to a desired yield;

f) allowing at least one of excess air and waste gases to leave said container continuously via said gas outlet means;

g) checking for at least one of contaminants and the quality of at least one of the cells and tissues which are produced in said container: if contaminants are found or at least one of the cells and tissues which are produced are of poor quality, the device and its contents are disposed of; if contaminants are not found, step h) is executed;

h) harvesting said desired portion of said medium containing at least one of cells and tissue, while leaving said remainder of medium containing at least one of cells and tissue in said container, wherein said remainder of medium serves as inoculant for a next culture/harvest cycle;

i) providing sterile at least one of said culture medium and said additives for the next culture/harvest cycle via said additive inlet means;

j) repeating steps d), e), f), g), h) and i) a plurality of times until in g) the said contaminants are found or at least one of the cells and tissues which are produced are of poor quality, whereupon the device and its contents are disposed of.

47. The method as claimed in claim 46, wherein said device further comprises air inlet means for introducing sterile air in the form of bubbles into said culture medium through a first inlet opening connectable to a suitable sterile air supply, said method further comprising the step of providing sterile air to said air inlet means during the first and each subsequent cycle.

48. The method as claimed in claim 47, wherein said sterile air is supplied continuously throughout at least one culturing cycle.

49. The method as claimed in claim 47, wherein said sterile air is supplied in pulses during at least one culturing cycle.

50. A method for axenically culturing and harvesting at least one of tissue and a quantity of cells in a battery of disposable devices comprising the steps of:

a) providing a battery of devices,

said battery comprising at least two devices, at least one of said devices comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissue and quantity of cells when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle, and

(iv) an air inlet means for introducing sterile air in the form of bubbles into said culture medium through a first inlet opening, wherein said air inlet means is connectable to a suitable air supply,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided, and

said harvesting means of each device is suitably connected to a common harvesting piping having a free end optionally comprising suitable aseptic connecting means thereat and said harvesting means further comprises a contamination prevention means for substantially preventing introduction of contaminants into said container via said common harvesting piping, said contamination prevention means comprising a U-shaped fluid trap, said fluid trap having one arm thereof free and further having an opening and wherein the other end of said contamination prevention means is aseptically mountable to said free end of said common harvesting piping via suitable aseptic connection means

and for at least one said device thereof:

b) providing axenic inoculant to said device via said common harvesting piping;

c) providing sterile at least one of said culture medium and said additives to said device via said common additive inlet piping;

k) optionally illuminating said device with external light means;

l) allowing at least one of said cells and tissue in said device to grow in said medium to a desired yield;

m) allowing at least one of excess air and waste gases to leave said device continuously via said common gas outlet piping;

n) checking for at least one of contaminants and the quality of at least one of the cells and tissues which are produced in said device: if in said device contaminants are found or at least one of the cells and tissues which are produced are of poor quality, said harvesting means of said device is closed off preventing contamination of other said devices of said battery; if in all of said devices of said battery contaminants are found or at least one of the cells and tissues which are produced therein are of poor quality, all the devices and their contents are disposed of; if contaminants are not found and the quality of at least one of the produced cells and tissues is acceptable, the device is considered harvestable and step h) is executed;

o) for each said harvestable device of step g), harvesting said desired portion of said medium containing at least one of cells and tissue via said common harvesting piping and said contamination prevention means to a suitable receiving tank, while leaving said remainder of medium containing at least one of said cells and tissue in said container, wherein said second portion of medium serves as inoculant for a next culture/harvest cycle;

p) providing sterile at least one of said culture medium and said additives for the next culture/harvest cycle via said additive inlet means;

q) repeating steps d), e), f), g), h) and i) a plurality of times until in g) said contaminants are found or at least one of the cells and tissues which are produced are of poor quality for all of said devices of said battery, whereupon said contamination prevention means are disconnected from said common harvesting means and said devices and their contents are disposed of.

51. A method for axenically culturing and harvesting at least one of tissue and a quantity of cells in a battery of disposable devices comprising the steps of:

a) providing a battery of devices,

said battery of devices comprising at least two disposable devices, at least one of said devices comprising a sterilisable disposable container which is at least one of transparent and translucent, having a top end and a bottom end, which container may be at least partially filled with at least one of a suitable sterile biological cell culture medium, a suitable sterile biological tissue culture medium, axenic inoculant, sterile air and required other sterile additives, said container comprising:

(i) gas outlet means for removing at least one of excess air and waste gases from said container;

(ii) additive inlet means for introducing at least one of said inoculant, said culture medium and said additives into said container;

and characterized in further comprising

(iii) at least one air inlet means for introducing sterile air in the form of bubbles into said culture medium through a first inlet opening, wherein said air inlet means is suitably connected to a common air inlet piping having a free end optionally comprising suitable aseptic connecting means thereat, which is further connectable to a suitable air supply;

(iv) reusable harvesting means comprising suitable flow control means for enabling harvesting of at least a desired portion of said medium containing at least one of said tissues and quantity of cells when desired, thereby enabling said device to be used continuously for at least one further consecutive culturing/harvesting cycle,

wherein a remainder of said medium containing at least one of said cells and tissue, remaining from a previous harvested cycle, may serve as inoculant for a next culture and harvest cycle, wherein at least one of said culture medium and said required additives are provided;

and for at least one said device thereof:

b) providing axenic inoculant to said device via said common harvesting piping;

c) providing sterile at least one of said culture medium and said additives to said device via said common additive inlet piping;

d) providing sterile air to said device via said common air inlet piping;

e) optionally illuminating said device with external light means;

f) allowing at least one of said cells and tissue in said device to grow in said medium to a desired yield;

g) allowing at least one of said excess air and waste gases to leave said device continuously via said common gas outlet piping;

h) checking for at least one of contaminants and the quality of at least one of the cells and tissues which are produced in said device: if in said device contaminants are found or at least one of the cells and tissues which are produced are of poor quality, said harvesting means of said device is closed off preventing contamination of other said devices of said battery; if in all of said devices of said battery contaminants are found or at least one of the cells and tissues which are produced therein are of poor quality, all the devices and their contents are disposed of; if contaminants are not found and the quality of at least one of the produced cells and tissues is acceptable, the device is considered harvestable and step i) is executed;

i) for each of said harvestable device of step h), harvesting at least a desired portion of said medium containing at least one of said cells and tissue via said common harvesting piping and said contamination prevention means to a suitable receiving tank, while leaving said remainder of medium containing at least one of the cells and tissue in said container, wherein said remainder of medium serves as inoculant for a next culture/harvest cycle;

j) providing sterile at least one of said culture medium and said additives for the next culture/harvest cycle via said additive inlet means;

k) repeating steps d), e), f), g), h), i) and j) a plurality of times until in h) said contaminants are found or at least one of the cells and tissues which are produced are of poor quality for all of said devices of said battery, whereupon said contamination prevention means are disconnected from said common harvesting means and said devices and their contents are disposed of.

52. The method as claimed in claim 46, wherein said remainder of said medium containing at least one of said cells and tissue comprises between about 2.5% and about 45% of the original volume of said culture medium and said inoculant.

53. The method as claimed in claim 52, wherein said remainder of said medium containing at least one of said cells and tissue comprises between about 10% and about 20% of the original volume of said culture medium and said inoculant.

54. The method as claimed in claim 47, wherein said step of providing sterile air further comprises the step of producing air bubbles, some of said air bubbles comprising a mean diameter of between about 1 mm and about 10 mm.

55. The method as claimed in claim 54, wherein at least some of said air bubbles comprise a mean diameter of about 4 mm.

56. The device as claimed in claim 31, wherein said container comprises an internal fillable volume of between about 50 liters and 150 liters.

57. The device as claimed in claim 31, wherein said container comprises an internal fillable volume of about 100 liters.
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