Bioreactor system

• US 10,240,121 B2
• Filed: 08/16/2011
• Issued: 03/26/2019
• Est. Priority Date: 08/16/2010
• Status: Active Grant

• Current Assignee: Kiyatec Inc.
• Original Assignee: Kiyatec Inc.
• Inventors: Gevaert, Matthew R., Orr, David E., Widener, Margaret Rebecca
• Primary Examiner(s): Humphrey, Louise W
• Assistant Examiner(s): Perkins, Stephen A
• Application Number:
  US13/817,636
• Publication Number:
  US 20130309771A1
• Time in Patent Office: 2,779 Days
• Field of Search: 422503, 4283044, 42831111, 42831171, 4283122, 4283126, 4353091
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 23/44 Multiple separable units; Modules
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture

Abstract

Disclosed herein are bioreactor systems and methods of utilizing said systems.

Claims

1. A bioreactor system for 3D passaging of cells comprising;

at least one cell module defining a first cell culture chamber, an inlet, an outlet, and a port opening, wherein the port opening is on one end of the cell culture chamber; and

a scaffold, located within the at least one cell module, comprising a stacked crosshatch of struts or fibers defining the pores of the scaffold, and a cell anchorage deposited within the pores of the scaffold, wherein cells attach to the cell anchorage.

2. The bioreactor system of claim 1, wherein the scaffold has a porosity between 50 μ

m and 2 mm.

3. The bioreactor system of claim 1, wherein the struts or fibers are made from metal, ceramic, glass, or polymers.

4. The bioreactor system of claim 1, wherein the cell anchorage comprises a polymer selected from the group consisting of polylactide, chondroitin sulfate, polyesters, polyethylene glycols, polycarbonates, polyvinyl alcohols, polyamides, polyacrylamides, poly methacrylate, polyether esters, polyurethanes, polycaprolactone, polvphosphazenes, polyorthoesters, polyglycolide, copolymers of lysine and lactic acid, copolymers of lysine-RGD lactic acid, chitosan, agarose, alginate, collagen, hyaluronic acid, carrageenan, demineralized bone matrix, or a combination thereof.

5. The bioreactor system of claim 4, further comprising a cell encapsulated in a hydrogel.

6. The bioreactor system of claim 1, wherein the cell anchorage is released from the scaffold by chemical degradation, light exposure, change in temperature, change in pH, or exposure to a chemical catalyst.

7. The bioreactor system of claim 1, wherein the cell anchorage is introduced into or onto the scaffold prior to being loaded into the bioreactor system.

8. The bioreactor system of claim 7, wherein a cell is encapsulated in the cell anchorage prior to introduction to the scaffold.

9. The bioreactor system of claim 1, wherein the cell anchorage is introduced into the scaffold after the scaffold is loaded into the bioreactor system.

18. The bioreactor system of claim 1, wherein the cell anchorage has a Young'"'"'s modulus of about 500 MPa or less.

10. A method of 3D cell passaging in a bioreactor system, comprising:

providing a population of cells to be passaged; and

introducing the population of cells into a bioreactor system comprising a scaffold, the scaffold comprising a stacked crosshatch of struts or fibers defining pores in the scaffold, and a cell anchorage deposited within the pores of the scaffold, wherein the cells attach to the cell anchorage.

11. The method of claim 10, wherein the cells are introduced into the cell anchorage prior to introduction of the cell anchorage to the struts or fibers.

12. The method of claim 10, further comprising causing the cell anchorage to disassociate from the stiff struts or fibers.

13. The method of claim 12, further comprising isolating the cells and cell anchorage from the bioreactor system.

14. The method of claim 12, further comprising recovering dissociated cells and dividing the cells into multiple populations.

15. The method of claim 14, further comprising introducing one or more divided cells into a second bioreactor system.

16. The method of claim 15, wherein the second bioreactor system comprises a second scaffold, the second scaffold comprising a stacked crosshatch of struts or fibers defining pores in the scaffold, and a cell anchorage that is deposited within the pores of the second scaffold.

17. The method of claim 16, wherein the cells are introduced into the cell anchorage prior to introduction of the cell anchorage into the pores of the second scaffold.