US2010248366A1PendingUtilityA1

Synthetic Surfaces for Differentiating Stem Cells into Cardiomyocytes

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Assignee: GERON CORPPriority: Jan 30, 2008Filed: Feb 8, 2010Published: Sep 30, 2010
Est. expiryJan 30, 2028(~1.6 yrs left)· nominal 20-yr term from priority
Inventors:Andrei Gennadyevich FadeevJennifer Lynn WeberZara MelkoumianDavid M. WeberYue ZhouRalph Brandenberger
C12N 5/0606C12N 2537/10C12N 2533/50C12N 2533/30C12N 5/0068C12N 5/0602
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Claims

Abstract

Synthetic surfaces capable of supporting culture of eukaryotic cells including stem cells and undifferentiated human embryonic stem cells in a chemically defined medium include a swellable (meth)acrylate layer and a polypeptide conjugated to the swellable (meth)acrylate layer. The swellable (meth)acrylate layer may be formed by polymerizing monomers in a composition that includes a carboxyl group-containing (meth)acrylate monomer, a cross-linking (di- or higher-functional) (meth)acrylate monomer, and a hydrophilic monomer capable of polymerizing with the carboxyl group-containing (meth)acrylate monomer and the cross-linking (meth)acrylate monomer. The swellable (meth)acrylate layer has an equilibrium water content in water of between about 5% and about 70%. The conjugated peptide may include an RGD amino acid sequence.

Claims

exact text as granted — not AI-modified
1 .- 25 . (canceled) 
     
     
         26 . A method of differentiating a pluripotent stem cell into a cardiomyocyte comprising culturing the pluripotent stem on a surface comprising a swellable (meth)acrylate layer conjugated to an RGD peptide and contacting the pluripotent stem cells with a media comprising one or more growth factors. 
     
     
         27 . The method of  claim 26 , wherein the pluripotent stem cell is a human embryonic stem cell. 
     
     
         28 . The method of  claim 26 , wherein the swellable (meth)acrylate layer comprises hydroxyethyl methacrylate. 
     
     
         29 . The method of  claim 26 , wherein the swellable (meth)acrylate layer comprises 2-carboxyethyl acrylate. 
     
     
         30 . The method of  claim 26 , wherein the swellable (meth)acrylate layer comprises tetra(ethylene glycol) dimethacrylate. 
     
     
         31 . The method of  claim 26 , wherein the swellable (meth)acrylate layer comprises hydroxyethyl methacrylate, 2-carboxyethyl acrylate, and tetra(ethylene glycol) dimethacrylate. 
     
     
         32 . The method of  claim 26 , wherein the growth factors are activin and BMP4. 
     
     
         33 . The method of  claim 32 , wherein the pluripotent stem cells are contacted with activin first and then BMP4. 
     
     
         34 . The method of  claim 26 , wherein the cardiomyocytes express Nkx2.5. 
     
     
         35 . The method of  claim 26 , wherein the cardiomyocytes express actinin. 
     
     
         36 . The method of  claim 26 , wherein the RGD peptide is a fragment of bone sialo protein. 
     
     
         37 . The method of  claim 26 , wherein the RGD peptide is a fragment of vitronectin.

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