Swellable (meth)acrylate surfaces for culturing cells in chemically defined media
Synthetic surfaces capable of supporting culture of undifferentiated human embryonic stem cells in a chemically defined medium include a swellable (meth)acrylate layer and a peptide conjugated to the swellable (meth)acrylate layer. The swellable (meth)acrylate layer may be formed by polymerizing monomers in a composition that includes hydroxyethyl methacrylate, 2-carboxyehylacrylate, and tetra(ethylene glycol) dimethacrylate. The conjugated peptide may include an amino acid sequence of Xaa n ProGlnValThrArgGlyAspValPheThrMetPro, where n is an integer from 0 to 3 and where Xaa is any amino acid. Further, disclosed herein is a swellable (meth)acrylate synthetic surface which can be sterilized by gamma irradiation.
1. A cell culture article comprising:
a swellable (meth)acrylate layer, wherein the swellable (meth)acrylate layer is formed from a composition comprising 2-carboxyethylacrylate, tetra(ethylene glycol) dimethacrylate, and one of hydroxyethyl methacrylate, hydroxypropyl methacrylate, 2-hydroxyethyl acrylate, 1-vinyl-2-pyrrolidone, di(ethylene glycol) ethyl ether methacrylate, acrylamide, ethylene glycol methyl ether methacrylate or 3-sulfopropyldimethyl-3-methacrylamidopropyl-ammonium; and
a peptide comprising an amino acid sequence of XaanProGlnValThrArgGlyAspValPheThrMetPro (SEQ ID NO:1), wherein Xaa is any amino acid and n is an integer from 0 to 3;
wherein the peptide is conjugated to the swellable (meth)acrylate layer; and,
wherein the cell culture article has been sterilized by gamma irradiation.
2. The cell culture article of claim 1 , wherein the swellable (meth)acrylate layer is free of polypeptide crosslinkers.
3. The cell culture article of claim 1 , wherein the swellable (meth)acrylate layer is formed from a composition comprising hydroxyethyl methacrylate, 2-carboxyethylacrylate, and tetra(ethylene glycol) dimethacrylate.
4. The cell culture article of claim 1 , wherein the swellable (meth)acrylate layer is formed from a composition comprising about 60-90% hydroxyethyl methacrylate by volume, about 10-40% 2-carboxyethyacrylate by volume, and about 1-5% tetra(ethylene glycol) dimethacrylate by volume.
5. The cell culture article of claim 1 , wherein the volume ratio of hydroxyethyl methacrylate/2-carboxyethylacrylate/tetra(ethylene glycol) dimethacrylate is about 80/20/3.
6. The cell culture article of claim 4 , wherein the swellable (meth)acrylate layer is formed from a composition consisting essentially of hydroxyethyl methacrylate/2-carboxyethylacrylate/tetra(ethylene glycol) dimethacrylate at a volume ratio of about 80/20/3.
7. The cell culture article of claim 3 , wherein the swellable (meth)acrylate layer is a uniform layer.
8. The cell culture article of claim 1 , wherein the polypeptide is conjugated to the swellable (meth)acrylate layer at a density of greater than about 5 picomoles per square millimeter of a surface of the swellable (meth)acrylate layer.
9. The cell culture article claim 1 wherein the gamma irradiation comprises 10-25 kGy treatment.
10. A cell culture system comprising:
a swellable (meth)acrylate layer, wherein the swellable (meth)acrylate layer is formed from a composition comprising 2-carboxyethylacrylate, tetra(ethylene glycol) dimethacrylate, and one of hydroxyethyl methacrylate, hydroxypropyl methacrylate, 2-hydroxyethyl acrylate, 1-vinyl-2-pyrrolidone, di(ethylene glycol) ethyl ether methacrylate, acrylamide, ethylene glycol methyl ether methacrylate or 3-sulfopropyldimethyl-3-methacrylamidopropyl-ammonium; and
a peptide comprising an amino acid sequence of Xaa n ProGlnValThrArgGlyAspValPheThrMetPro (SEQ ID NO:1), wherein n is an integer from 0 to 3 and wherein Xaa is any amino acid;
wherein the peptide is conjugated to the swellable (meth)acrylate layer;
a chemically defined medium; and
one or more undifferentiated human stem cells, wherein the cells are in the medium and are in contact with the peptide.
11. The system of claim 10 , wherein the chemically defined medium comprises basic fibroblast growth factor and transforming growth factor-β1.
12. A method for culturing undifferentiated stem cells, comprising:
contacting the undifferentiated human stem cells with the peptide conjugated swellable (meth)acrylate layer of the article of claim 1 ; and
culturing the cells in a cell culture medium to maintain the cell in an undifferentiated state.
13. The method of claim 12 , wherein the culture medium is a chemically defined culture medium.
14. The method of claim 13 , wherein the culture medium comprises basic fibroblast growth factor and transforming growth factor-β1.
15. The method of claim 14 , wherein the culture medium comprises about 80 ng/ml basic fibroblast growth factor and about 0.5 ng/ml transforming growth factor-β1.