IP Library Granted Patent US 12,378,517
Granted Patent B2
US 12,378,517 · App. 18/656,253 · Granted Aug 5, 2025

Perfusion culturing methods and uses thereof

Inventors: Agata Villiger-Oberbek (Cambridge, MA); Jianguo Yang (Sudbury, MA); Yang Yang (Hopkinton, MA); Gabrielle Lorenzo (West Seneca, NY)
Assignee: Genzyme Corporation
C12N5/0602C12M23/12C12M27/10C12M29/10C12P21/00C12N2510/02C12N2527/00
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Quick Facts
Patent No.
US 12,378,517
App. No.
18/656,253
Granted
Aug 5, 2025
Kind
B2
Abstract

Provided herein are methods of culturing a mammalian cell and various methods that utilize these culturing methods. Also provided are multi-well cell culture plates, e.g., for use in perfusion culturing methods.

Claims (25)

1. A multi-well cell culture plate system, comprising:

a unitary support plate comprising a first surface comprising a plurality of apertures;

a plurality of culture vessels disposed within the unitary support plate and configured to house cell cultures having a volume of between about 200 μL to about 18 mL in volume, wherein each aperture is paired with and defines an opening into each culture vessel and wherein each culture vessel further comprises at least one port configured to accommodate a flow of fluid into and/or out of the culture vessel, each port in fluid connection with its own conduit, each conduit disposed within the unitary support plate and defines its own fluid flow passage connecting the port and the first surface.

2. The multi-well cell culture plate system of claim 1 , wherein each culture vessel comprises at least a first port and a second port, wherein the first port is configured to accommodate a one-way flow of fluid into the culture vessel and the second port is configured to accommodate a one-way flow of fluid out of the culture vessel.

3. The multi-well cell culture plate system of claim 1 , wherein a port of the at least one port comprises a filter configured to selectively prevent cells from flowing into and out of the culture vessel.

4. The multi-well cell culture plate system of claim 2 , wherein the first port and the second port each comprise a filter configured to selectively prevent cells from flowing into and out of the culture vessel.

5. The multi-well cell culture plate system of claim 1 , further comprising at least one fluid flow regulator operably connected to the at least one port.

6. The multi-well cell culture plate system of claim 1 , further comprising at least one fluid flow meter operably connected to the at least one port.

7. The multi-well cell culture plate system of claim 1 , wherein the plurality of culture vessels are configured to house cell cultures having a volume of between about 1 mL to about 10 mL in volume.

8. The multi-well cell culture plate system of claim 7 , wherein the plurality of culture vessels are configured to house cell cultures having a volume of between about 1 mL to about 5 mL in volume.

9. The multi-well cell culture plate system of claim 1 , wherein the plurality of culture vessels each have a cylinder shape with an end opposite to that of the aperture that is flat, hemispherical, pyramidal, or conical.

10. The multi-well cell culture plate system of claim 1 , wherein the at least one port is a one-way valve.

11. The multi-well cell culture plate system of claim 1 , wherein the at least one port is a two-way valve.

12. The multi-well cell culture plate system of claim 1 , wherein each of the plurality of culture vessels is a square-bottom well.

13. The multi-well cell culture plate system of claim 12 , wherein each of the plurality of culture vessels has a diameter of between about 6.0 mm and about 35 mm.

14. The multi-well cell culture plate system of claim 12 , wherein each of the plurality of culture vessels has a height of between about 40 mm and about 50 mm.

15. The multi-well cell culture plate system of claim 1 , wherein the plurality of culture vessels is 6, 12, 24, 48, or 96 culture vessels.

16. The multi-well cell culture plate system of claim 1 , wherein each of the plurality of culture vessels are sealed with a gas-permeable disposable membrane.

17. The multi-well cell culture plate system of claim 1 , wherein each of the plurality of culture vessels are sealed with a gas-permeable silicone layer.

18. A multi-well cell culture plate system, comprising:

a unitary support plate comprising a first surface comprising a plurality of apertures;

a reservoir within the unitary support plate configured to store a fluid, wherein the reservoir comprises an exchange port configured to receive the fluid from a surface of the unitary support plate that is different than the first surface;

a plurality of culture vessels disposed within the unitary support plate and configured to house cell cultures having a volume of between about 200 μL to about 18 mL in volume, wherein each aperture is paired with and defines an opening into each culture vessel and wherein each culture vessel further comprises at least one port configured to accommodate a flow of the fluid between the culture vessel and the reservoir.

19. The multi-well cell culture plate system of claim 18 , wherein the exchange port is a two-way valve and is arranged on a side surface of the unitary support plate.

20. The multi-well cell culture plate system of claim 18 , wherein a port of the at least one port comprises a filter configured to selectively prevent cells from flowing into and out of the culture vessel.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 3, 2024
From: VILLIGER-OBERBEK, AGATA; YANG, YANG; LORENZO, GABRIELLE; YANG, JIANGUO
To: GENZYME CORPORATION
Reel/Frame 067904/0139 →
Continuity (4)
Continuation 17358889 · Jun 25, 2021
Continuation 14732325 · Jun 5, 2015
Provisional Application 62009058 · Jun 6, 2014
Related Publication 20240287451A1 · Aug 29, 2024
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