Defined surfaces of self-assembled monolayers and stem cells
A method for the construction of arrays from self-assembling monolayers is described. The arrays have particular utility for the screening of peptides ligands that can foster the growth of cells in culture. This technique has been used to identify peptide ligands that foster the growth of human stem cells, which otherwise require an extracellular matrix in order to grow in an undifferentiated state. This also makes possible an assay to identify other such peptides.
1. A human stem cell culture comprising:
human stem cells, a medium in which human stem cells will grow, and a self-assembled monolayer presenting to the human stem cells molecules of a peptide ligand that supports the growth of human stem cells in an undifferentiated state, wherein the peptide ligand consists essentially of a receptor recognition epitope, wherein the monolayer comprises a plurality of elements, each element comprising an attachment moiety, an alkane spacer moiety attached to the attachment moiety, and a peptide ligand covalently attached to the spacer moiety such that all peptide ligands within an array element are presented on the array in a uniform orientation, the monolayer arranged on an inert substrate with the attachment moiety of each element attached to the substrate.
2. A human stem cell culture as claimed in claim 1 , wherein the peptide ligand is selected from the group consisting of SDPGYIGSR (SEQ ID NO:2), GRNIAEIIKDI (SEQ ID NO:3), DITYVRLKF (SEQ ID NO:4), DITVTLNRL (SEQ ID NO:5), GRYVVLPR (SEQ ID NO:6), GNRWHSIYITRFG (SEQ ID NO:7), SIDQVEPYSSTAQ (SEQ ID NO:8), KIPKASSVPTELSAISTLYL (SEQ ID NO:9), KKQRFRHRNRKG (SEQ ID NO:10), GASIKVAVSADR (SEQ ID NO:11), GTTVKYIFR (SEQ ID NO:12), GSIKIRGTYS (SEQ ID NO:13) and GSINNNR (SEQ ID NO:14).
3. A self-assembled monolayer presenting molecules of a peptide ligand that support the growth of human stem cells in an undifferentiated state, wherein the peptide ligand consists essentially of a receptor recognition epitope, wherein the monolayer comprises a plurality of elements, each element comprising an attachment moiety, an alkane spacer moiety attached to the attachment moiety, and a peptide ligand covalently attached to the spacer moiety such that all peptide ligands within an array element are presented on the array in a uniform orientation, the monolayer arranged on an inert substrate with the attachment moiety of each element attached to the substrate.
4. A self-assembled monolayer as claimed in claim 3 , wherein the peptide ligand is selected from the group consisting of SDPGYIGSR (SEQ ID NO:2), GRNIAEIIKDI (SEQ ID NO:3), DITYVRLKF (SEQ ID NO:4), DITVTLNRL (SEQ ID NO:5), GRYVVLPR (SEQ ID NO:6), GNRWHSIYITRFG (SEQ ID NO:7), SIDQVEPYSSTAQ (SEQ ID NO:8), KIPKASSVPTELSAISTLYL (SEQ ID NO:9), KKQRFRHRNRKG (SEQ ID NO:10), GASIKVAVSADR (SEQ ID NO:11), GTTVKYIFR (SEQ ID NO:12), GSIKIRGTYS (SEQ ID NO:13) and GSINNNR (SEQ ID NO:14).
5. A method for culturing undifferentiated human stem cells, the method comprising the steps of:
applying the stem cells and a medium in which human stem cells will grow to a self-assembled monolayer presenting to the human stem cells molecules of a peptide ligand that support the growth of human stem cells in an undifferentiated state, wherein the monolayer comprises a plurality of elements, each element comprising an attachment moiety, an alkane spacer moiety attached to the attachment moiety, and a peptide ligand covalently attached to the spacer moiety such that all peptide ligands within an array element are presented on the array in a uniform orientation, the monolayer arranged on an inert substrate with the attachment moiety of each element attached to the substrate, wherein the peptide ligand consists essentially of a receptor recognition epitope; and
culturing the cells under conditions that favor the growth of cells.
6. A method as claimed in claim 5 , wherein the peptide ligand is selected from the group consisting of SDPGYIGSR (SEQ ID NO:2), GRNIAEIIKDI (SEQ ID NO:3), DITYVRLKF (SEQ ID NO:4), DITVTLNRL (SEQ ID NO:5), GRYVVLPR (SEQ ID NO:6), GNRWHSIYITRFG (SEQ ID NO:7), SIDQVEPYSSTAQ (SEQ ID NO:8), KIPKASSVPTELSAISTLYL (SEQ ID NO:9), KKQRFRHRNRKG (SEQ ID NO:10), GASIKVAVSADR (SEQ ID NO:11), GTTVKYIFR (SEQ ID NO:12), GSIKIRGTYS (SEQ ID NO:13) and GSINNNR (SEQ ID NO:14).