Cell culture media and methods

Patent metadata

Field	Value
Publication number	US-12203095-B2
Application number	US-202016918838-A
Country	US
Kind code	B2
Filing date	Jul 1, 2020
Priority date	Dec 22, 2011
Publication date	Jan 21, 2025
Grant date	Jan 21, 2025

Abstract

Compositions and methods are described for preparing media, feeds, and supplements. Such methods and medias may display increased stability of labile components and may use, for example, microsuspension and/or encapsulation technologies, chelation, and optionally, coating and/or mixing the labile compounds with anti-oxidants. The compositions may withstand thermal and/or irradiation treatment and have reduced virus number. These techniques may result in product with extended shelf-life, extended release of their internal components into culture, or in product that can be added aseptically into a bioreactor using minimal volumes. The compositions and methods may optimize the bioproduction workflow and increase efficiency.

First claim

What is claimed is:

1. A serum-free cell culture medium composition comprising a microsuspension comprising a mixture of a labile component and an antioxidant that is microencapsulated into a bead with a capsular matrix, wherein the bead has a diameter within the range of 0.05-1.5 mm, wherein a powdered cell culture medium is used for the microencapsulated microsuspension, wherein the composition is irradiated with gamma-rays, and wherein the labile component is embedded in the anti-oxidant to protect the labile component by reducing the impact of oxidation species generated during irradiation.

2. The serum-free cell culture medium composition of claim 1 , wherein the powdered cell culture medium is an advanced granulation technology (AGT) cell culture medium.

3. The serum-free cell culture medium composition of claim 1 , wherein the material of the capsular matrix is selected from the group consisting of alginate, poly-L-lactic acid, chitosan, agarose, gelatin, hyaluronic acid, chondroitin sulfate, dextran, dextran sulfate, heparin, heparin sulfate, heparan sulfate, gellan gum, xanthan gum, guar gum, water soluble cellulose derivatives and carrageenan.

4. The serum-free cell culture medium composition of claim 1 , wherein the capsular material is soluble upon reconstitution with an aqueous solvent.

5. The serum-free cell culture medium composition of claim 1 , wherein the bead is coated with a coating solution.

6. The serum-free cell culture medium composition of claim 5 , wherein the coating solution is selected form the group consisting of poly-glycolic acid, PLGA (poly-lactic-co-glycolic acid), collagen, polyhydroxyalkanoates (PHA), poly-ε-caprolactone, poly-ortho esters, poly-anhydrides, poly-phosphazenes, poly-amino acids, polydimethylsiloxane, polyurethranes, poly-tetrafluoroethylene, polyethylene, polysulphone, poly-methyl methacrylate, poly-2-hydroxyethylmethacrylate, polyamides, polypropylene, poly-vinyl chloride, polystyrene, poly-vinyl pyrrolidone, poly-L-lysine and polyornithine.

7. The serum-free cell culture medium composition of claim 1 , wherein the bead is additionally irradiated with UV rays.

8. The serum-free cell culture medium composition of claim 7 , wherein the bead is free of PPV and MMV viruses.

9. The serum-free cell culture medium composition of claim 1 , wherein the gamma-rays are 25-100 kGy or 30-50 kGy.

10. The serum-free cell culture medium composition of claim 1 , wherein the composition is protein free.

11. The serum-free cell culture medium composition of claim 1 , wherein the composition is a dry-format media.

12. The serum-free cell culture medium composition of claim 1 , wherein the labile component is selected from the group consisting of a polyamine, a growth factor, a cytokine and a vitamin.

13. The serum-free cell culture medium composition of claim 1 , wherein the composition further comprises chelated reactive species and said chelated reactive species are selected from the group consisting of cations, metals ions or trace elements.

14. The serum-free cell culture medium composition of claim 1 , wherein the composition further comprises chelating moieties and said chelating moieties are selected from the group consisting of EDTA, citrate, succinate, cyclodextrin, clatharates, dendrimers and amino acids.

15. The serum-free cell culture medium composition of claim 1 , wherein the composition is added aseptically into a bioreactor.

Assignees

• Life Technologies Corp

Inventors

• Fike Richard
• Barrett Shawn
• Jiang Zhou
• Judd David

Classifications

• C12N2500/38

  Vitamins · CPC title

• B01J13/22

  Coating · CPC title

• B01J13/06

  by phase separation · CPC title

• C12N2533/74

  Alginate · CPC title

• C12N2533/40

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