pH ADJUSTMENT TO IMPROVE THAW RECOVERY OF CELL BANKS

a cell bank and thaw recovery technology, applied in the field of freezing cells, can solve problems such as inconsistent or poor viability, and achieve the effect of improving the thaw recovery of cell banks

Inactive Publication Date: 2020-10-08
GENENTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent text describes a method for improving the recovery of cells that have been frozen and stored in a medium containing a buffered solution and a cryoprotective agent. The method involves adjusting the pH of the medium to a value between 6.7 and 8.5 before freezing the cells. This results in better cell survival and recovery after thawing. The technical effect of this method is to provide a more effective and reliable way to freeze and store eukaryotic cells.

Problems solved by technology

These traditional methods may result in inconsistent or poor viability post-thaw for select cell lines.

Method used

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  • pH ADJUSTMENT TO IMPROVE THAW RECOVERY OF CELL BANKS
  • pH ADJUSTMENT TO IMPROVE THAW RECOVERY OF CELL BANKS
  • pH ADJUSTMENT TO IMPROVE THAW RECOVERY OF CELL BANKS

Examples

Experimental program
Comparison scheme
Effect test

example 1

Pool pH, Hold Temperature and Hold Time on Thawed Cell Viability

[0102]Base Addition Method to Control Banking pH (Small Scale Study #1)

[0103]A study was executed to gauge the feasibility of performing a simple pH adjustment from worst case and typical pool conditions (targeted initial pH of 6.2 and 6.7 respectively) to enhance thaw recovery. Several pH adjustment targets were studied to determine optimal banking pH. The pH was measured using the on BioProfile® 400 (Nova Biomedical) instrument with the temperature set to 37° C. Similarly, pH calculations using Equation 1 are based upon a sample temperature of 37° C.

[0104]Cells were pelleted via centrifugation. Cell pools were generated for each test condition by resuspending cells to 100×106 cells / mL in spent media. The cell pools were agitated at 37° C. with or without gas permeable membranes for CO2 / O2 exchange to drive pH to initial pH conditions (either 6.2 or 6.7). Upon reaching initial pH targets, pools were quickly chilled to ...

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Abstract

Provided herein are methods of freezing mammalian cells for storage or improving thaw recovery of cell banks comprising freezing mammalian cells in a freezing medium having a pH of 6.7 to 8.5.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the priority benefit of U.S. provisional application Ser. No. 62 / 022,392, filed Jul. 9, 2014, which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION[0002]The present invention relates to methods of freezing cells (such as mammalian cells) for banking and freezing media for use in freezing cells.BACKGROUND OF THE INVENTION[0003]Cell banks are produced by first accumulating cells in a batch / perfusion cell culture process and then harvesting cells for banking. The process involves three stages: cell accumulation, harvest and cell concentration, and cell banking. A harvest process step serves to concentrate the final cell culture fluid or to extract the cells from the cell culture fluid using centrifugation. A subsequent pooling and filling process serves to prepare cell bank ampoules for long-term storage.[0004]These traditional methods may result in inconsistent or poor viability post-thaw f...

Claims

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Application Information

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Patent Type & AuthorityApplications(United States)
IPC IPC(8): A01N1/02C12N5/00C12N5/071
CPCC12N2510/02C12N5/0018A01N1/0221C12N5/0682A01N1/0284A01N1/0278
InventorMEIER, ANGELAMEIER, STEVEN J.DUFFY, PHILLIPCOYNE, MARCIACALHOUN, KARA
OwnerGENENTECH INC