CELL-CULTURE-BAG

• US 20130157353A1
• Filed: 05/11/2011
• Published: 06/20/2013
• Est. Priority Date: 05/12/2010
• Status: Abandoned Application

• Current Assignee: Xpand Biotechnology BV (Kuros Biosciences Ltd.)
• Original Assignee: Xpand Biotechnology BV (Kuros Biosciences Ltd.)
• Inventors: Dijkhuizen Borgart, Elise Leonore Isolde, Bracke, Madelon Sophia George Maria, De Bruijn, Joost Dick
• Application Number:
  US13/643,282
• Publication Number:
  US 20130157353A1
• Time in Patent Office: Days
• US Class Current: 435/297.2
• CPC Class Codes: C12M 23/14 Bags
• CPC Class Codes: C12M 25/16 Particles; Beads; Granular material; Encapsulation chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00
• CPC Class Codes: C12M 27/16 Vibrating; Shaking; Tilting
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 41/44 of volume or liquid level

Abstract

A cell-culture-bag for use in the expansion of stem cells from a crude biopsy, comprising: outer walls; a chamber located within the walls; a first inlet, and first and second outlets, providing fluid communication with the chamber, for connection to a perfusion apparatus; and a filter arrangement constructed to allow passage of red blood cells and block passage of stem cells from the first outlet, and block passage of microcarriers from the second outlet.

Claims

1. A cell-culture-bag for use in the expansion of stem cells from a crude biopsy, comprising:

outer walls;

a chamber located within the walls;

a first inlet, and first and second outlets, providing fluid communication with the chamber, for connection to a perfusion apparatus; and

a filter arrangement constructed to allow passage of red blood cells and block passage of stem cells from the first outlet, and block passage of microcarriers from the second outlet.

2. A cell-culture-bag as in claim 1, wherein the filter arrangement filters the fluid path from the chamber to the first outlet with a mesh of 8-20 μ

m.

3. A cell-culture-bag as in claim 1, wherein the filter arrangement filters the fluid path from the chamber to the second outlet with at least a mesh of 50-100 μ

m.

4. A cell-culture-bag as in claim 1, further comprising a third outlet, and a filter arrangement being constructed to block the passage of at least microcarriers from the third outlet.

5. A cell-culture-bag as in claim 4, wherein the filter arrangement filters the fluid path from the chamber to the third outlet with a mesh of 50-100 μ

m.

6. A cell-culture-bag as in claim 1, wherein the filter arrangement comprises an inner filter bag and an outer filter bag that encloses the inner filter bag.

7. A cell-culture-bag as in claim 1, wherein the filter arrangement comprises a filter member that is associated with a said outlet and joins to the portion of the outlet wall surrounding the said outlet so as to sealingly enclose said outlet.

8. A cell-culture-bag as in claim 7, wherein the filter member has an area which is substantially greater than that of the associated outlet.

9. A cell-culture-bag as in claim 7, wherein the join between the filter member and the outer wall includes a protuberance that creates a small clearance space between the filter member and the outer wall.

10. A cell-culture-bag as in claim 7, wherein each of said outlets has a respective said filter member.

11. A cell-culture-bag as in claim 1, wherein each of said outlets and the inlet has an associated coupling for connection to a conduit of the perfusion apparatus.

12. A cell-culture-bag as in claim 1, which is constructed so as to permit the partitioning of the chamber into sub-chambers.

13. A system for expanding stem cells from crude biopsy, comprising:

a cell-culture-bag according to claim 1; and

a perfusion apparatus connected to the first inlet, and outlets for performing seeding, expansion, harvesting and collection operations on stem cells placed within the chamber.

14. A system as in claim 13, further comprising a means for partitioning the chamber into a first sub-chamber and a second sub-chamber, wherein the partitioning means allows the relative volume of the first and second sub-chambers to be adjusted.

16. The cell-culture bag of claim 4 wherein said third outlet blocks the passage of stem cells from said outlet.

17. The cell-culture bag of claim 16 wherein the filter arrangement filters the fluid path from the chamber to the third outlet with a mesh of 8-20 μ

m.

18. The system of claim 14 wherein the partitioning means allows the relative volume of the first and second sub-chambers to be adjusted.

15. A system for expanding cells from a crude biopsy, comprising:

a cell-culture-bag comprising a chamber;

a first inlet, and a seeding/expansion outlet, and a collection outlet, each in fluid communication with the chamber;

a perfusion apparatus connected to the first inlet, and the seeding/expansion and collection outlets;

wherein the perfusion apparatus has a seeding mode in which the first inlet and the seeding/expansion outlet are in operative connection with a seeding circuit of the perfusion apparatus, and a seeding operation is performed on stem cells within the chamber;

wherein the perfusion apparatus has an expansion mode in which the first inlet and the seeding/expansion outlet are in operative connection with an expansion circuit of the perfusion apparatus, and an expansion operation is performed on stem cells within the chamber;

wherein the perfusion apparatus has a harvesting mode in which the first inlet is in operative connection with a harvesting section of the perfusion apparatus, and an harvesting operation is performed on stem cells within the chamber;

wherein the perfusion apparatus has a collection mode in which the collection outlet is in operative connection with a collection section of the perfusion apparatus, and harvested stem cells exit from the collection outlet.