Biomaterials for 3D Cell Growth and Differentiation
AssigneeTexas Tech University System
InventorsHarvinder Singh Gill, Chang Hyun Lee
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Abstract
The present invention includes a polypeptide for use in a three dimensional (3D) culture system for the growth of cells comprising one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X1 and X2 are any amino acids except proline, wherein X1 and X2 can be the same or different amino acid in solution or coated on a substrate, wherein n1 and n2 are equal to or greater than one, and wherein X is an aliphatic amino acid.
Claims28
A polypeptide for use in a three dimensional (3D) culture system for the growth of cells comprising:
- one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X, X1, and X2 can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one.
A nucleic acid that encodes a polypeptide that comprises one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X, X1, and X2 can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one.
A nucleic acid vector that encodes a polypeptide that comprises one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X, X1, and X2 can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one;
- or the polypeptide has the sequence selected from at least one of [(X1X2GXP)(X3X4GXP)]n1, [(X1X2GXP)n1(X3X4GXP)n2], or [(X1X2GXP)n1(X3X4GXP)n2(X1X2GXP)n1], wherein X, X1, X2, X3, and X4 are any amino acid, and n1 and n2 are greater than or equal to one;
- wherein X1 and X2 can be the same or different from each other, and X3 and X4 can be the same or different from each other;
- or wherein at least one of X1 or X2 is different from X3 or X4, or wherein X is valine, or X1=G, X2=Y and A (in 1:4 ratio) and X=V.
A host cell that comprises a nucleic acid vector that encodes a polypeptide that comprises one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X, X1, and X2 can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one, and optionally the host cell expresses or secretes the polypeptide.
A method of making a fusion protein comprising:
- providing a host cell with a nucleic acid vector that expresses a polypeptide that comprises one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X, X1, and X2 can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one;
- and
- isolating the polypeptide.
A method of making cardiomyocytes comprising:
- seeding stem cells and incubating in a media that comprise a polypeptide that comprises one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X1, X2 and X can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one;
- culturing the stem cells without an anti-differentiation factor;
- changing the media to cardiac differentiation media;
- and
- isolating beating cardiomyocytes.
A beating cardiomyocyte made by a method comprising:
- seeding stem cells in a media comprising a polypeptide that comprises one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X1, X2 and X can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one;
- culturing the stem cells without an anti-differentiation factor;
- changing the media to cardiac differentiation media;
- and
- isolating beating cardiomyocytes.
The method of claim 26, further comprising making a 3D cell culture comprising:
- seeding cells and incubating in a media that comprises a polypeptide that comprises one or more repeats of a sequence n1-(X1X2GXP)-n2 (SEQ ID NO:8), wherein X, X1, X2 are any amino acid, wherein X1, X2 and X can be the same or different amino acid, wherein n1 and n2 are equal to or greater than one;
- culturing cells;
- changing the media;
- and
- isolating the cells.
Dependencies are inferred from the claim text.