Cell Culture Media Supplement and Method of Molecular Stress Control

Abstract

A novel class of agents has been identified to serve as cell-guard agents and / or target-specific supplements to increase cell quality and yield, as well as select for target cell populations. Laboratory experiments have demonstrated the use of cell-guard agents and / or target-specific supplements in the bioprocessing of cells as well as in selecting out a desired cell population. Several potential additive agents (both natural and synthetic) have been identified during these studies, including Vitamin D3, NAC, resveratrol, salubrinal, AKT, and tunicamycin (among others) that hold promise for application in cell models. In one embodiment, hypothermic stress regimes are utilized. In another embodiment, normothermic conditions are utilized while other stressors are tested in the processing. The methods of maintaining mass cell cultures and / or selecting out particular cell populations for further research and clinical use represents an important step in therapeutic discovery. The cell-guard agents are process-matched and cell-matched to protect and / or select particular cells during laboratory and clinical manipulation. The components modulate cell stress and survival pathways.

Description

RELATED APPLICATIONS[0001]The present application claims priority to U.S. Provisional Patent Application Ser. No. 61 / 368,287 filed on Jul. 28, 2010 and titled Cell Culture Media Supplement and Method of Molecular Stress Control, which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION[0002]The present invention relates generally to the area of cell and molecular biology and, in particular, to supplements for use in bioprocessing.BACKGROUND OF THE INVENTION[0003]The utilization of in vitro cultured cells in research, medicine, bioprocessing, and bioproduction settings is expanding exponentially. While widely applied and often viewed as common practice, cell culture is a highly variable and dynamic process which has significant impact on the overall quality and performance of the culture. The nuances of cell culture are often addressed with simple procedural or pattern alterations in a laboratory setting, yet these alterations are more often than not incompatib...

AI Technical Summary

Benefits of technology

[0013]This invention utilizes unique compound classes to confer protection to cells undergoing bioprocessing events such as harvesting, bulk culture, fluorescence activated cell sorting, shipping, transfection and protein bioproduction. The cell culture media supplements, known as cell-guard and cell-select agents, improve the yield and function of human cells undergoing bioprocessing.

[0015]As a vitamin or therapeutic adjuvant to cell culture systems, cell-guard can be either a liquid supplement or dissolved tablet, proportional to the cell culture media undergoing various processes. In another aspect, RNAi may be utilized to augment cell-guard function and enhance cell quality. The cell-guard additive reduces the level of cellular loss associated with cell therapy applications and improves viability of media compositions.

[0016]In addition, the control of a molecular pathway can also be moderated by using inhibitors to turn on / off a response pathway (e.g. UPR, apoptosis, heat shock, etc.). This mechanism is observed in the use of such inhibitors as caspase, AKT, MPTP, and salubrinal in modulating pathways to improve usability of human cell types. Key cell stress pathways have been identified to target the use of cell-guard additives for growth of hepatocytes, human pancreatic islets, mesenchymal stem cells, cancer stem cells, human corneal endothelial cells (HCEC), hepatoma cell lines (C3A) and kidney cell lines (786-O), among others.

[0019]The overall intent of the cell-guard technology is to maintain cells in a more native, reduced stress response molecular-based disposition during bioprocessing. In one aspect, one cell-guard compound enables cells to be maintained in vitro in a more “normal functional state” for extended periods, thereby extending usable life and performance. The approach to developing the cell-guard technology is based on an understanding of the molecular response mechanisms activated within a cell in response to various stressors. It is through an investigation of the cell's stress response that cell-guard is capable of being an additive or supplement compatible with various culture media and bioreactors so that cell stress can be reduced, thus reducing apoptosis and increasing the production (usable) life of existing bioprocessing cultures.

Problems solved by technology

The nuances of cell culture are often addressed with simple procedural or pattern alterations in a laboratory setting, yet these alterations are more often than not incompatible with scale up for successful batch culture of cells in bioprocessing settings.

Mass production and utilization of cell media offers a number of technical challenges starting with the simple fact that cell culture is typically characterized by low growth and production rates in comparison with chemical processes.

While cellular bioprocessing has become integral in basic and applied research, as well as in medical therapeutics, a number of significant obstacles have resulted from the stresses created by the conditions used to support proper cell growth and function in an in vitro environment.

As such, much of the classical cell culture media formulation fundamentals fall short in providing a means to support efficient cell bioprocessing.

In this regard, challenges remain to develop a specialized culture media that can be customized to individual cell types and culture production processes, procedures and protocols, and the individual response of cells to variances in each of the other parameters associated with cell culture.

Further, in the field of medical diagnostics, including stem cell therapy and cancer, a challenge exists to eliminate or select one or more cell types from a mixed population of cancerous and non-tumorigenic cells without the use of fluorescent tags, chemotherapeutic agents or antibodies.

Also, the ability to selectively eliminate cancer cells in a cell culture setting using mutagenic, chemotherapeutic compounds is the basis of cancer drug discovery; but this approach typically requires the use of DNA binding drugs and / or tumor suppressor activators that cannot be used in any procedure that ultimately results in the transfusion of cell products into patients.

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