Polymeric carriers and methods

Patent metadata

Field	Value
Publication number	US-11560545-B2
Application number	US-201816610325-A
Country	US
Kind code	B2
Filing date	May 1, 2018
Priority date	May 2, 2017
Publication date	Jan 24, 2023
Grant date	Jan 24, 2023

Abstract

Provided are methods of controlling disassociation of cells from a carrier, compositions, and methods of collecting cells. The methods of controlling disassociation of cells from a carrier may include contacting a polymeric carrier with one or more digesting agents to disassociate at least a portion of a plurality of cells from the polymeric carrier. The polymeric carrier may be crosslinked with a crosslinker including at least one of a redox sensitive moiety, a UV light sensitive moiety, a pH sensitive moiety, and a temperature sensitive moiety.

First claim

The invention claimed is:

1. A method of controlling disassociation of cells from a carrier, the method comprising:
providing a polymeric carrier and a plurality of cells adhered to the polymeric carrier, wherein the polymeric carrier is a microcarrier; and
contacting the polymeric carrier with one or more digesting agents to disassociate at least a portion of the plurality of cells from the polymeric carrier;
wherein the polymeric carrier comprises gelatin, and is crosslinked with a crosslinker comprising at least one redox sensitive moiety, wherein the at least one redox sensitive moiety comprises a disulfide bond, and the crosslinker comprises a compound having the following structure—

wherein R 1 and R 2 are independently selected from a divalent C 1 -C 20 hydrocarbyl.

2. The method of claim 1 , wherein R 1 and R 2 are unsubstituted divalent C 1 -C 5 hydrocarbyls, and the crosslinker has the following structure:

3. The method of claim 1 , wherein R 1 and R 2 are unsubstituted divalent C 2 hydrocarbyls, and the crosslinker comprises 1,2-bis-(2-isocyanatoethyl)disulfide (DIDS):

4. The method of claim 1 , wherein the plurality of cells comprises human Mesenchymal stem cells (hMSCs), endothelial cells, or a combination thereof.

5. The method of claim 1 , wherein the polymeric carrier further comprises dextran, hyaluronic acid, agarose, chitosan, alginate, polyvinyl alcohol, polyamino acid, calcium alginate, polyacrylate, polyethylene glycol, polyethylene imine, polyanhydride, or a combination thereof.

6. The method of claim 1 , wherein the polymeric carrier further comprises dextran, hyaluronic acid, agarose, chitosan, alginate, polyvinyl alcohol, polyamino acid, or a combination thereof.

7. The method of claim 1 , wherein the polymeric carrier is a polymeric carrier bead.

8. The method of claim 1 , wherein the one or more digesting agents comprises a reducing agent.

9. The method of claim 8 , wherein the reducing agent comprises dithiothreitol (DTT), cysteine, glutathione, or a combination thereof.

10. The method of claim 8 , wherein the reducing agent comprises a solution of dithiothreitol (DTT) having a concentration of about 5 mM to about 25 mM.

11. The method of claim 10 , wherein the solution of dithiothreitol further comprises sodium citrate, trypsin, or a combination thereof.

12. The method of claim 1 , wherein the one or more digesting agents comprises an enzyme.

13. The method of claim 12 , wherein the enzyme comprises collagenase, dispase-like protease, trypsin, urease, papain, bromelain, pancreatin, gelatinase (MMP2 and MMP9), or a combination thereof.

14. The method of claim 1 , wherein the one or more digesting agents comprises a reducing agent and an enzyme.

15. The method of claim 1 , wherein the ratio of the crosslinker (mmol) to the polymeric carrier (g) is about 0.1 mmol:10 g to about 1.9 mmol:10 g.

Assignees

• Brigham & Womens Hospital Inc
• Lonza Walkersville Inc
• Massachusetts Inst Technology

Inventors

• Zhang Yi
• Artzi Natalie
• Wang Kui
• Abraham Eytan
• Levinson Yonatan

Classifications

• C08L5/08

  Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof · CPC title

• C08L5/04

  Alginic acid; Derivatives thereof · CPC title

• C12N2533/40

  Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers · CPC title

• C12N2537/10

  Cross-linking · CPC title

• C08L2312/00

  Crosslinking · CPC title