Chemically labile peptide-presenting surfaces for cellular self-assembly
Methods of cell culture using patterned SAM arrays are disclosed. Advantageously, the disclosed methods use SAM arrays presenting adhesion peptides to grow confluent monolayers that can invaginate to form an embryoid body.
1. A method of controlling the formation of a cell culture aggregate, the method comprising:
forming on a substrate, at least one alkanethiolate self-assembled monolayer spot wherein the spot is conjugated to a cellular adhesive peptide consisting of SEQ ID NO: 4 using a labile covalent bond, wherein the spot is part of a self-assembled monolayer array, and wherein the self-assembled monolayer array is prepared using a method selected from the group consisting of microcontact printing, microfluidics, stamping, photochemistry, locally removing a region in a fully formed self-assembled monolayer and reforming a new self-assembled monolayer in the region;
culturing at least one cell on the alkanethiolate self-assembled monolayer spot for a sufficient time to form a confluent monolayer of cells; and
detaching the confluent monolayer of cells from the array spot by latent nucleophilic cleavage of the labile covalent bond between the cellular adhesive peptide and the alkanethiolate, wherein the detached confluent monolayer of cells forms the cell culture aggregate; and
collecting the cell culture aggregate.
2. The method of claim 1 , wherein the confluent monolayer is cultured for a period of from about 6 hours to about 144 hours.
3. The method of claim 1 , further comprising culturing the confluent monolayer for a sufficient time to allow the confluent monolayer to invaginate.
4. The method of claim 3 , wherein the confluent monolayer is cultured for a period of from about 6 hours to about 144 hours.
5. The method of claim 1 , wherein the cell is selected from the group consisting of an induced pluripotent stem cell, a mesenchymal stem cell, an umbilical vein endothelial cell, a dermal fibroblast, a fibrosarcoma cell, an embryonic stem cell, an iPS IMR90-4 cell, an iPS-derived endothelial cell, and combinations thereof.
6. The method of claim 1 , wherein the specified diameter of the array spot is from about 600 μm to about 6 mm.
7. The method of claim 1 , wherein said aggregates comprises either a uniform size or a specified shape.
8. The method of claim 7 , wherein the specified shape is selected from the group consisting of a circle, an oval, and oval cross, a star, and a hand.