Claims for Patent: 5,622,857

Summary for Patent: 5,622,857

Title:	High performance cell culture bioreactor and method
Abstract:	A high performance hollow fiber bioreactor having concentric hollow fiber bundles: a central hollow fiber bundle supplies media, and an outer array supplies oxygen needed for cell culture. Useful to expand therapeutic cells such as stem cells ex vivo, and as an extracorporeal device such as an artificial liver.
Inventor(s):	Goffe; Randal A. (Arlington, WA)
Assignee:	Genespan Corporation (Redmond, WA)
Application Number:	08/512,546
Patent Claims:	1. A bioreactor comprising: a housing comprising an inner wall defining a substantially tubular cell culture chamber, first and second ends defining a longitudinal axis through said chamber, and a port providing access to said chamber, a central bundle of porous hollow fibers disposed around the longitudinal axis within said chamber, an annular bundle of gas permeable hollow fibers disposed concentric to and surrounding said central bundle, means disposed at said first and second ends for passing nutrient media through said porous hollow fibers, and means disposed at said first and second ends for passing an oxygen containing gas through said gas permeable hollow fibers. 2. The bioreactor of claim 1, wherein an annular space is provided between said inner wall and said annular bundle. 3. The bioreactor of claim 2, further comprising a plurality of microcarriers within said annular space. 4. The bioreactor of claim 1, wherein an annular space is provided between said annular bundle and said central bundle. 5. The bioreactor of claim 4, further comprising a plurality of microcarriers within said annular space. 6. The bioreactor of claim 1, wherein the ratio of the surface area of said gas permeable hollow fibers to the surface area of said porous hollow fibers is from about 0.5 to about 3.1, and wherein said gas permeable hollow fibers and said porous hollow fibers collectively occupy from about 10 to about 40 percent by volume of the cell culture chamber. 7. The bioreactor of claim 6, wherein the ratio of the surface area of said gas permeable hollow fibers to the surface area of said porous hollow fibers is from about 0.5 to about 1.5, and wherein said gas permeable hollow fibers and said porous hollow fibers collectively occupy from about 10 to about 25 percent by volume of the cell culture chamber. 8. The bioreactor of claim 6, wherein the ratio of the surface area of said gas permeable hollow fibers to the surface area of said porous hollow fibers is from about 1.5 to about 3.1, and wherein said gas permeable hollow fibers and said porous hollow fibers collectively occupy from about 25 to about 40 percent by volume of the cell culture chamber. 9. The bioreactor of claim 1, wherein the ratio of the surface area of said gas permeable hollow fibers to the surface area of said porous hollow fibers is from about 3.1 to about 4, and wherein said gas permeable hollow fibers and said porous hollow fibers collectively occupy from about 40 to about 50 percent by volume of the cell culture chamber. 10. The bioreactor of claim 1, wherein the ratio of the surface area of said gas permeable hollow fibers to the surface area of said porous hollow fibers is from about 4 to about 5.8, and wherein said gas permeable hollow fibers and said porous hollow fibers collectively occupy from about 50 to about 70 percent by volume of the cell culture chamber. 11. A kit comprising a bioreactor of claim 2 and a plurality of microcarriers. 12. The kit of claim 11, wherein said microcarriers are coated with an extracellular matrix. 13. A kit comprising a bioreactor of claim 6 and a plurality of microcarriers. 14. A kit comprising a bioreactor of claim 9 and a plurality of microcarriers. 15. A kit comprising a bioreactor of claim 4 and a plurality of microcarriers. 16. The kit of claim 15, wherein said microcarriers are coated with an extracellular matrix. 17. A method of cell culture comprising: introducing cells into the cell culture chamber of a bioreactor of claim 1, and incubating said cells within said chamber while passing an oxygen containing gas through said gas permeable hollow fibers and while passing nutrient media through said porous hollow fibers. 18. The method of claim 17, wherein the passage of oxygen containing gas through said gas permeable hollow fibers is counter current to the passage of nutrient media through said porous hollow fibers. 19. The method of claim 17 wherein during said incubation step the bioreactor is periodically rotated to-and-fro around its longitudinal axis. 20. The method of claim 17, wherein said nutrient media contains a free radical scavenger. 21. The method of claim 20, wherein said free radical scavenger is selected from among the group consisting of sodium benzoate and benzoic acid. 22. The method of claim 20, wherein said free radical scavenger is present in said nutrient media at a concentration of from about 0.001 to about 0.1 M. 23. The method of claim 22, wherein said nutrient media containing said free radical scavenger is buffered to maintain pH at about 7.1. 24. The method of claim 17, wherein said oxygen containing gas is passed through said gas permeable hollow fibers at a pressure low enough to avoid formation of gas bubbles within the cell culture chamber. 25. The method of claim 24, wherein said pressure is from about 0.5 to about 1.5 psi. 26. The method of claim 17, wherein said bioreactor comprises an annular space between said inner wall and said annular bundle, and wherein said cells are introduced through said port into said annular space. 27. The method of claim 26, wherein a plurality of microcarriers are provided in said annular space. 28. The method of claim 17, wherein said bioreactor comprises an annular space between said annular bundle and said central bundle, and wherein said cells are introduced through said port into said annular space. 29. The method of claim 28, wherein a plurality of microcarriers are provided in said annular space. 30. The method of claim 17, further comprising: harvesting said cells from said chamber. 31. The method of claim 30, wherein said bioreactor comprises a first port adjacent said first end and a second port adjacent said second end, and wherein said harvesting step comprises introducing a gas through the first port into the chamber and thereby displacing medium containing cells from the chamber through the second port. 32. The method of claim30, wherein said harvesting step comprises introducing a proteolytic enzyme into the chamber. 33. The method of claim 32, wherein the proteolytic enzyme is selected from among the group consisting of trypsin and collagenase. 34. A bioreactor comprising: a housing comprising an inner wall defining a substantially tubular cell culture chamber, the chamber having first and second ends, a longitudinal axis, and a port providing access to said chamber; a central bundle of porous hollow fibers surrounding the longitudinal axis of said chamber, each of the fibers in fluid communication with a common axially-extending liquid inlet port; and an annular bundle of gas permeable hollow fibers disposed concentric with, and surrounding, said central bundle, each of the gas permeable fibers of the annular bundle having an outlet end and an inlet end, the outlet ends of the gas permeable fibers in fluid communication with a common annular space surrounding the axially-extending liquid inlet port, a gas outlet port in fluid communication with the common annular space. 35. The bioreactor of claim 34, wherein the annular bundle is spaced from the inner wall of the cell culture chamber to provide a cell culture annular space. 36. The bioreactor of claim 35, further comprising a plurality of microcarriers within said cell culture annular space. 37. The bioreactor of claim 34, wherein the ratio of the surface area of said gas permeable hollow fibers to the surface area of said porous hollow fibers is from about 0.05 to about 3.1. 38. The bioreactor of claim 37, wherein the central bundle of porous hollow fibers and the annular bundle of gas permeable fibers collectively occupy about 10 to about 40% by volume of the chamber. 39. The bioreactor of claim 34, wherein the ratio of the surface area of the gas permeable fibers to the hollow fibers is from about 0.5 to about 1.5 40. The bioreactor of claim 39, wherein the annular bundles of gas permeable hollow fibers and the central bundle of porous hollow fibers occupy collectively from about 10 to about 25% by volume of the cell culture chamber. 41. The bioreactor of claim 34, wherein the ratio of the surface area of said gas permeable hollow fibers to the surface area of said porous hollow fibers is from about 1.5 to about 3.1. 42. The bioreactor of claim 41, wherein the central bundle and the annular bundle collectively occupy from about 25 to about 40% of the volume of the cell culture chamber. 43. The bioreactor of claim 34, wherein the ratio of the surface area of the gas permeable hollow fibers to the porous hollow fibers is from about 4 to about 5.8. 44. The bioreactor of claim 43, wherein the gas permeable fibers and the porous fibers collectively occupy from about 50 to about 70% by volume of the cell culture chamber. 45. The bioreactor of claim 34, wherein the central bundle is spaced from the annular bundle. 46. The bioreactor of claim 35, wherein the central bundle is spaced from the annular bundle.

Details for Patent 5,622,857

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2014-02-09