System for in-situ management of pH of culture

Patent metadata

Field	Value
Publication number	US-9677046-B2
Application number	US-201314418880-A
Country	US
Kind code	B2
Filing date	Jul 31, 2013
Priority date	Jul 31, 2012
Publication date	Jun 13, 2017
Grant date	Jun 13, 2017

Abstract

Disclosed is a hydrogel composition loaded with a base that regulates and maintains the pH of cell culture media. The invention particularly discloses in situ pH managing hydrogels (pHmH) loaded with base Mg(OH) 2 for pH control in small scale cultures comprising adherent mammalian cells, suspended mammalian cells, microorganisms, a microorganism with plasmids and the like thereof.

First claim

What claimed is:

1. A system for in situ management of pH of a culture comprising a carrier and a base, wherein said base is Mg(OH) 2 or Ca(OH) 2 , and said carrier is selected from the group consisting of a hydrogel, a polymer matrix comprising a silicone elastomer, an encapsulation for said base, a membrane and a coating film,
wherein release rate of said base is dependent on pH of the culture, said carrier is non-responsive to pH change of said culture, and wherein the carrier is loaded with the base.

2. The system of claim 1 , wherein the carrier is a hydrogel.

3. The system of claim 2 , wherein the hydrogel comprises 2-hydroxyethyl methacrylate (HEMA) as monomer; Azoisobutyronitrile, (AIBN) as initiator and optionally ethylene glycol dimethacrylate (EGDMA) as cross linker.

4. The system of claim 3 , wherein an amount of the HEMA is from 71.8 to 99.9% by weight of total amount of the hydrogel.

5. The system of claim 4 , wherein the base is loaded in the carrier in a manner selected from the group consisting of:
the base is dispersed in the carrier, and
the base is included in a reservoir within the carrier.

6. The system of claim 1 , wherein said culture comprises medium and cells selected from the group consisting of unmodified animal cells, genetically modified animal cells, microbial cells, insect cells, stem cells and mesenchymal stem cells.

7. The system of claim 1 , wherein said culture is grown in a medium comprising glucose, and optionally a buffer.

8. The system of claim 1 , wherein the pH of the culture is managed between 5-9.

9. The system of claim 1 , wherein the pH of the culture is managed for a period of up to 15 days.

10. The system of claim 1 , wherein Mg(OH) 2 or Ca(OH) 2 is released depending on pH of the culture.

11. The system of claim 1 , wherein Mg(OH) 2 or Ca(OH) 2 is released depending on pH of the culture, and wherein said pH dependent-release of Mg(OH) 2 or Ca(OH) 2 is independent of the carrier.

12. The system of claim 1 , wherein the in situ management does not require periodic measurement of pH.

13. The system of claim 1 , wherein the system comprises two discs of the carrier.

14. A process for in situ management of pH of a culture, the process comprising employing a system comprising;
a carrier selected from the group consisting of a hydrogel, a polymer matrix comprising a silicone elastomer, an encapsulation for said base, a membrane and a coating film, wherein said carrier is non-responsive to pH change of said culture; and
a base, wherein said base is Mg(OH) 2 or Ca(OH) 2 , wherein release rate of said base is dependent on pH of the culture, and the carrier is loaded with the base.

Assignees

• Council Scient Ind Res

Inventors

• Gadgil Mugdha Chetan

Classifications

• C12N2500/34

  Sugars · CPC title

• C12N2500/16

  Magnesium; Mg chelators · CPC title

• C12N2533/30

  Synthetic polymers · CPC title

• C12N5/0068 Primary

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title