Thermoresponsive cell culture supports

Patent metadata

Field	Value
Publication number	US-9701939-B2
Application number	US-201214110465-A
Country	US
Kind code	B2
Filing date	Apr 6, 2012
Priority date	Apr 8, 2011
Publication date	Jul 11, 2017
Grant date	Jul 11, 2017

Abstract

The present invention relates to a cell culture support comprising a substrate and a thermoresponsive polymeric blend layer, wherein the polymeric blend layer comprises at least one thermoresponsive polymer and at least one network forming adhesion promoter. The present invention further relates a method of making a cell culture complex comprising: providing a substrate; blending at least one thermoresponsive polymer and at least one network forming adhesion promoter to provide a polymeric blend; applying a thin film of said polymeric blend to the substrate to provide a polymeric blend layer on the substrate; curing the polymeric blend layer on the substrate to provide a cell culture support; and depositing cells onto said cell culture support, wherein the cells may optionally further comprise medium, to provide a cell culture complex.

First claim

What is claimed is:

1. A cell culture support comprising:
a substrate;
a polymeric blend layer comprising at least one thermoresponsive polymer and at least one network forming adhesion promoter, characterized in that said network forming adhesion promoter is in the form of a cross-linked network bound to the substrate and said cross-linked network interlocks chains of the thermoresponsive polymer; and wherein the network forming adhesion promoter is a non-protein adhesion promoter and has functional amino or carboxylic acid groups; and
cells supported by said polymeric blend layer, wherein said thermoresponsive polymer provides for temperature induced detachment of said cells.

2. The support of claim 1 , wherein the substrate is selected from the group consisting of polymeric materials, glasses, ceramics, metals, metal oxides, hydrated metal oxides, and combinations thereof.

3. The support of claim 1 , wherein the at least one thermoresponsive polymer is selected from the group consisting of poly (N-isopropylacrylamide) (PNIPAAm), poly(N,N-diethylacrylamide) (PDEAAm), poly(N-vinlycaprolactam) (PVCL), poly[2-(dimethylamino) ethyl methacrylate] (PDMAEMA), and poly(ethylene oxide) (PEO), and combinations thereof.

4. The support of claim 1 , wherein the at least one thermoresponsive polymer is poly (N-isopropylacrylamide) represented by the formula:

5. The support of claim 1 , wherein the at least one network forming adhesion promoter is an aminosilane.

6. The support of claim 1 , wherein the at least one network forming adhesion promoter is selected from the group consisting of 3-aminopropyltriethoxysilane (APTES), 3-aminopropyldiethoxymethylsilane (APDEMS), and 3-aminopropyltrimethoxysilane (APTMS), and combinations thereof.

7. The support of claim 1 , wherein the at least one network forming adhesion promoter is 3-aminopropyltriethoxysilane (APTES) represented by the formula:

8. The support of claim 1 , wherein the polymer blend layer is characterized as having a thermoresponsive polymer to network forming adhesion promoter ratio (TRP:NFAP) of from about 99:1 to about 40:60 by mass, and wherein the polymer blend layer is cured to produce a film.

9. The support of claim 1 , wherein the polymeric blend layer is devoid of cell adhesive proteins.

10. The support of claim 1 , wherein the polymeric blend layer is devoid of plasma or e-beam treatment.

11. The support of claim 1 , wherein the support supports said cells such that said temperature induced detachment is achieved without mechanical agitation or enzymatic aided detachment methods.

Assignees

• Newby Bi-Min
• Patel Nikul
• Cavicchia John
• Zhang Ge
• Univ Akron

Inventors

• Newby Bi-Min
• Patel Nikul
• Cavicchia John
• Zhang Ge

Classifications

• C12N5/0662

  Stem cells · CPC title

• C12N5/0664

  Dental pulp stem cells, Dental follicle stem cells · CPC title

• C12N5/0668

  Mesenchymal stem cells from other natural sources · CPC title

• C12N5/0666

  Mesenchymal stem cells from hair follicles · CPC title

• C12N5/0068 Primary

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