Thermoresponsive cell culture supports

Abstract

The present invention relates to a cell culture support comprising a substrate and a polymeric blend layer bound to the substrate. The polymeric blend layer comprises at least one thermoresponsive polymer and at least one coupling agent. The coupling agent is a non-protein coupling agent that has functional thiol, ester, epoxy, or aldehyde groups. The cell culture support further includes cells supported by the polymeric blend layer, wherein the thermoresponsive polymer provides for temperature induced detachment of the cells and / or cell sheets.

Description

RELATED APPLICATIONS[0001]This application claims the priority of provisional U.S. Application Ser. No. 62 / 341,703 filed May 26, 2016.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This invention was made with government support under NIH 1R15GM097626-01A1 awarded by National Institutes of Health. The government has certain rights in the invention.TECHNICAL FIELD[0003]The present invention relates to cell culture supports and methods of making cell culture supports. More particularly, this invention relates to thermoresponsive polymeric cell culture supports that are receptive to cell attachment and subsequent rapid cell sheet detachment. These supports are suitable for use, for example, in biomedical applications such as tissue engineering.BACKGROUND OF THE INVENTION[0004]Previous methods of preparing thermo-responsive cell culture supports have used two primary approaches, namely electron beam irradiation or plasma polymerization, to covalently graft poly (N-...

AI Technical Summary

Benefits of technology

The present invention is related to a cell culture support for growing anchor dependent cells, which can be used for regenerative medicine purposes. The cell culture support can be patterned and assembled to mimic the microarchitecture of native tissue. Additionally, the cells to be cultured can be autologous or heterozoic, and they can be used for various applications such as fibroblasts, myoblasts, and myotube cells. The use of human mesenchymal stem cells (hMSCs) is also mentioned. The invention allows for increased cell-cell interactions and eliminates the risk of immunogenic materials present in scaffolds. The creation of the cell culture support involves depositing cells onto a polymeric blend layer and growing them to create the cultured cell sheet. The cells are then detached and used for various applications.

Problems solved by technology

The complicated procedures and apparatus required in these methods prevent a cost-effective adoption of this technology for specific applications.

Furthermore, previous methods have generally required the use of added adhesive proteins from other individuals or other species to enhance cell attachment, and such foreign additions tend to cause immunogenic reactions of the cell sheet products after transplantation.

Other methods that do not employ thermo-responsive polymers (TRPs) for detachment and harvesting of cell sheets from cell culture supports have employed proteolytic enzymes (e.g. trypsin) or mechanical agitations, which have resulted in damage to cells and their excreted extracellular matrix (ECM), thus negatively affecting their biological functions.

However, these methods are still too expensive for wide-spread use.

As previously mentioned, added adhesive proteins are undesirable as they may deleteriously cause immunogenic reactions.

However, the dissolution of poly-N-isopropylacrylamide into the cell culture medium upon cooling is undesirable.

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