Synthetic composition and coating for cell culture
A composition for forming a polymeric cell culture surface includes (i) a pre-polymer comprising a polymer backbone, a cationic moiety conjugated to the backbone, and a cross-linker moiety conjugated to the backbone; and (ii) a peptide-polymer comprising a polymer backbone and cell adhesive peptide conjugated to the backbone. Cross-linked coatings for cell culture that have a suitable amount of cell adhesive peptide and cationic moiety may be formed from the pre-polymer and peptide-polymer.
1. A composition for forming a polymeric cell culture surface comprising:
a pre-polymer comprising a polymer backbone, an amine cationic moiety conjugated to the backbone wherein the amine cationic moiety results from polymerization of a monoalkylaminoalkyl (meth) acrylate, N-tert-butylaminoethyl (meth)acrylate, or a dialkylaminoalkyl (meth)acrylate monomer selected from the group consisting of N,N-dimethylaminoethyl(meth)acrylate, N—N-diethylaminoethyl (meth) acrylate and N,N-dipropylaminoethyl (meth) acrylate, and a cross-linker moiety conjugated to the backbone; and
a peptide-polymer comprising a polymer backbone and cell adhesive peptide conjugated to the backbone.
2. The composition of claim 1 , wherein the amine cationic moiety conjugated to the polymer backbone results from the polymerization of a monomer into the pre-polymer, wherein the monomer is selected from the group consisting of an N,N-dimethylaminoethyl(meth)acrylate, an N,N-diethylaminoethyl (meth)acrylate, and an N,N-dipropylaminoethyl (meth)acrylate.
3. The composition of claim 1 , wherein the amine cationic moiety conjugated to the polymer backbone results from polymerization of a 2-(dimethylamino) ethyl methacrylate (DMEMA) monomer to form the pre-polymer.
4. The composition of claim 1 , wherein the pre-polymer further comprises a hydrophilic moiety conjugated to the polymer backbone of the pre-polymer.
5. The composition of claim 4 , wherein the hydrophilic moiety conjugated to the polymer backbone results from polymerization of a monomer into the pre-polymer, wherein the monomer is selected from the group consisting of 2-carboxyethyl methacrylate, 2-carboxyethyl acrylate, acrylic acid, methacrylic acid, hydroxypropyl methacrylate, 2-hydroxyethyl methacrylate (HEMA), 2-hydroxyethyl acrylate, glycerol methacrylate, hydroxypropyl acrylate, 4-hydroxybutyl acrylate, 2-carboxyethyl acrylamide, acrylamidoglycolic acid, N-(hydroxymethyl)acrylamide, N-[Tris(hydroxymethyl)methyl]acrylamide, 3-acryloylamino-1-propanol, N-acrylamido-ethoxyethanol, and N-hydroxyethyl acrylamide.
6. The composition of claim 4 , wherein the hydrophilic moiety conjugated to the polymer backbone results from polymerization of a 2-hydroxyethyl methacrylate (HEMA) monomer to form the pre-polymer.
7. The composition of claim 1 , wherein the cross-linker moiety is a benzophenone moiety.
8. The composition of claim 1 , wherein the cell adhesive peptide comprises an R-G-D sequence.
9. The composition of claim 1 , wherein the cell adhesive peptide is a sequence of amino acids found in vitronectin, laminin, bone sialoprotein, collagen, or fibronectin.
10. The composition of claim 1 , wherein the cell adhesive peptide comprises a sequence of amino acids selected from the group consisting of: KGGGQKCIVQTTSWSQCSKS (SEQ ID NO:1), GGGQKCIVQTTSWSQCSKS (SEQ ID NO:2), KYGLALERKDHSG (SEQ ID NO:3), YGLALERKDHSG (SEQ ID NO:4), KGGSINNNRWHSIYITRFGNMGS (SEQ ID NO:5), GSINNNRWHSIYITRFGNMGS (SEQ ID NO:6), KGGTWYKIAFQRNRK (SEQ ID NO:7), GGTWYKIAFQRNRK (SEQ ID NO:8), KGGTSIKIRGTYSER (SEQ ID NO:9), GGTSIKIRGTYSER (SEQ ID NO:10), KYGTDIRVTLNRLNTF (SEQ ID NO:11), YGTDIRVTLNRLNTF (SEQ ID NO:12), KYGSETTVKYIFRLHE (SEQ ID NO:13), YGSETTVKYIFRLHE (SEQ ID NO:14), KYGKAFDITYVRLKF (SEQ ID NO:15), YGKAFDITYVRLKF (SEQ ID NO:16), KYGAASIKVAVSADR (SEQ ID NO:17), YGAASIKVAVSADR (SEQ ID NO:18), CGGNGEPRGDTYRAY (SEQ ID NO:19), GGNGEPRGDTYRAY (SEQ ID NO:20), CGGNGEPRGDTRAY (SEQ ID NO:21), GGNGEPRGDTRAY (SEQ ID NO:22), KYGRKRLQVQLSIRT (SEQ ID NO:23), YGRKRLQVQLSIRT (SEQ ID NO:24), KGGRNIAEIIKDI (SEQ ID NO:25), GGRNIAEIIKDI (SEQ ID NO:26), KGGPQVTRGDVFTMP (SEQ ID NO:27), GGPQVTRGDVFTMP (SEQ ID NO:28), GRGDSPK (SEQ ID NO:29), KGGAVTGRGDSPASS (SEQ ID NO:30), GGAVTGRGDSPASS (SEQ ID NO:31), Yaa 1 PQVTRGNVFTMP (SEQ ID NO:32), RGDYK (SEQ ID NO:33), and combinations thereof.
11. The composition of claim 1 , wherein the cell adhesive peptide is incorporated into the peptide-polymer from a peptide monomer of the formula R m —S p —C ap ;
wherein R is a polymerization moiety selected from the group consisting of acrylate, methacrylate, acrylamide, methacrylamide, maleimide and fumarate and any combination thereof, and m is an integer greater than 1;
wherein S p is an optional spacer moiety wherein the spacer moiety comprises polyethylene oxide or polypropylene oxide having the formula (O—CH 2 CHR′) m2 where R′ is H or CH 3 and m2 is an integer from 0 to 20, or Xaa □ wherein Xaa is independently any amino acid and n is an integer from 0 to 20, or any combination thereof; and,
wherein C ap is a peptide comprising an R-G-D sequence.
12. The composition of claim 1 , wherein the peptide-polymer further comprises a hydrophilic moiety conjugated to the backbone of the peptide-polymer.
13. The composition of claim 12 , wherein the hydrophilic moiety conjugated to the polymer backbone results from polymerization of a monomer into the pre-polymer, wherein the monomer is selected from the group consisting of 2-carboxyethyl methacrylate, 2-carboxyethyl acrylate, acrylic acid, methacrylic acid, hydroxypropyl methacrylate, 2-hydroxyethyl methacrylate (HEMA), 2-hydroxyethyl acrylate, glycerol methacrylate, hydroxypropyl acrylate, 4-hydroxybutyl acrylate, 2-carboxyethyl acrylamide, acrylamidoglycolic acid, N-(hydroxymethyl)acrylamide, N-[Tris(hydroxymethyl)methyl]acrylamide, 3-acryloylamino-1-propanol, N-acrylamido-ethoxyethanol, and N-hydroxyethyl acrylamide.
14. The composition of claim 1 , wherein the peptide-polymer comprises the polymerization product of an ethylenically unsaturated monomer having a carboxyl group with an ethylenically unsaturated monomer not having a carboxyl group, wherein the cell adhesive peptide is conjugated to the pre-polymer via the carboxyl group, and wherein the ethylenically unsaturated monomer not having the carboxyl group is hydrophilic.
15. A method of making a cell culture article comprising providing the pre-polymer and peptide-polymer of claim 1 to a substrate and exposing the pre-polymer and peptide-polymer on the substrate to an energy source to cross-link the pre-polymer and peptide-polymer to each other and to the substrate.
16. A cell culture article made by the method of claim 15 .