Dendritic cell generator

Patent metadata

Field	Value
Publication number	US-12157901-B2
Application number	US-202218061732-A
Country	US
Kind code	B2
Filing date	Dec 5, 2022
Priority date	Jun 29, 2015
Publication date	Dec 3, 2024
Grant date	Dec 3, 2024

Abstract

Devices, systems, and methods can be used for the automated production of dendritic cells (DC) from dendritic cell progenitors, such as monocytes obtained from peripheral blood. The invention makes it possible to obtain sufficient quantities of a subject's own DC for use in preparing and characterizing vaccines, for activating and characterizing the activation state of the subject's immune response, and to aid in preventing and/or treating cancer or infectious disease.

First claim

What is claimed is:

1. A dendritic cell generation system comprising:
one or more dendritic cell differentiation cassettes comprising:
a plastic body comprising a cell culture chamber, the chamber comprising a monocyte-binding substrate;
a fluid inlet port; and
a fluid outlet port;
wherein the plastic body, the fluid inlet port, and the fluid outlet port are fluidically coupled to the cell culture chamber and configured to provide flow of a liquid culture medium across the substrate from the inlet port to the outlet port;

a culture medium reservoir fluidically coupled to the fluid inlet port of the one or more dendritic cell differentiation cassettes;
a fluid collection reservoir fluidically coupled to the fluid outlet port of the one or more dendritic cell differentiation cassettes; and
a pump configured for pumping fluid from the culture medium reservoir, through the cell culture chamber of the one or more dendritic cell differentiation cassettes, and into the fluid collection reservoir.

2. The dendritic cell generation system of claim 1 , wherein the pump mechanism comprises a pump built into each of the one or more dendritic cell differentiation cassettes.

3. The dendritic cell generation system of claim 1 , wherein the pump mechanism comprises one or more pumps detached from the one or more dendritic cell differentiation cassettes.

4. The dendritic cell generation system of claim 1 comprising a processor, a memory, and a set of instructions executable by the processor for carrying out a method of preparing dendritic cells of a subject.

5. The dendritic cell generation system of claim 1 that is configured as a single disposable unit.

6. The dendritic cell generation system of claim 1 that is configured for operation within a cell culture incubator.

7. The dendritic cell differentiation cassette of claim 1 , wherein the monocyte-binding substrate comprises a polystyrene surface that binds monocytes but not differentiated dendritic cells.

8. The dendritic cell differentiation cassette of claim 1 , wherein the monocyte-binding substrate forms a bottom of the cell culture chamber and comprises a flat surface.

9. The dendritic cell differentiation cassette of claim 1 , wherein the fluid inlet port and fluid outlet port are disposed at opposite ends of the cell culture chamber.

10. The dendritic cell differentiation cassette of claim 1 that is assembled from a polystyrene bottom slab and a poly(methyl methacrylate) (PMMA) top slab, the top and bottom slabs separated by a PMMA cell culture chamber slab.

11. A method of preparing dendritic cells of a subject, the method comprising the steps of:
providing a liquid sample comprising monocytes of the subject to a dendritic cell generation system comprising:
one or more dendritic cell differentiation cassette comprising:
a plastic body comprising a cell culture chamber, the chamber comprising a monocyte-binding substrate;
a fluid inlet port; and
a fluid outlet port;

wherein the plastic body, the fluid inlet port, and the fluid outlet port are fluidically coupled to the cell culture chamber and configured to provide flow of a liquid culture medium across the substrate from the inlet port to the outlet port;
a culture medium reservoir fluidically coupled to the fluid inlet port of the one or more dendritic cell differentiation cassettes;
a fluid collection reservoir fluidically coupled to the fluid outlet port of the one or more dendritic cell differentiation cassettes; and
a pump configured for pumping fluid from the culture medium reservoir, through the cell culture chamber of the one or more dendritic cell differentiation cassettes, and into the fluid collection reservoir;

pumping the liquid sample into the cell culture chamber of a dendritic cell differentiation cassette of the system, whereby at least a portion of said monocytes bind to the monocyte binding substrate of the cell culture chamber; and
pumping a dendritic cell differentiation medium into the cell culture chamber, whereby at least a portion of the bound monocytes differentiate into dendritic cells and detach from the monocyte-binding substrate.

12. The method of claim 11 , wherein the method further comprises pumping a dendritic cell maturation medium into the cell culture chamber to replace the dendritic cell differentiation medium, whereby at least a portion of the differentiated dendritic cells mature into mature dendritic cells.

13. The method of claim 11 , wherein the method further comprises collecting mature dendritic cells from the cell culture chamber.

14. The method of claim 13 , further comprising exposing the mature dendritic cells to one or more antigens.

15. The method of claim 11 , wherein the mature dendritic cells have the phenotype Lin- and CD4+.

16. The method of claim 11 , wherein the dendritic cell differentiation medium comprises IL4 and GM-CSF.

17. The method of claim 11 , wherein said dendritic cell differentiation medium is pumped through the cell culture chamber continuously for about 2-6 days at a flow rate from about 1 μL/min to about 10 μL/min.

18. The method of claim 11 , wherein the dendritic cell maturation medium comprises IL1β, IL6, TNFa, and PGE2.

19. The method of claim 18 , wherein said dendritic cell maturation medium is pumped through the cell culture chamber continuously for about 1-3 days at a flow rate from about 1 μL/min to about 10 μL/min.

20. The method of claim 11 , wherein dendritic cells from two or more subjects are prepared simultaneously.

Assignees

• Univ Northeastern

Inventors

• Murthy Shashi K
• Collier Bradley B

Classifications

• C12M25/02 Primary

  Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00) · CPC title

• C12N5/0639 Primary

  Dendritic cells, e.g. Langherhans cells in the epidermis · CPC title

• C12N5/0018

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title

• C12M1/22

  Petri dishes · CPC title

• C07K14/545

  IL-1 · CPC title