Synthetic poly
mer
microarray technology holds re
markable pro
mise to rapidly identify suitable bio
materials for ste
m cell and tissue engineering applications. However,
most of previous
microarrayed synthetic poly
mers do not possess biological ligands (e.g., peptides) to directly engage cell surface receptors. Here, we report the develop
ment of peptide-functionalized hydrogel
microarrays based on light-assisted copoly
merization of poly(ethylene glycol) diacrylates (PEGDA) and
methacrylated-peptides. Using solid-phase peptide/organic synthesis, we developed an efficient route to synthesize
methacrylated-peptides. In parallel, we identified PEG hydrogels that effectively inhibit non-specific cell adhesion by using PEGDA-700 (M. W. = 700) as a
mono
mer. The co
mbined use of these che
mistries enables the develop
ment of a powerful platfor
m to prepare peptide-functionalized PEG hydrogel
microarrays. Additionally, we identified a linker co
mposed of 4 glycines to ensure sufficient exposure of the peptide
moieties fro
m hydrogel surfaces. Further, we used this syste
m to directly co
mpare cell adhesion abilities of several related RGD peptides:
m>RGDm>,
m>RGDm>S,
m>RGDm>SG and
m>RGDm>SP. Finally, we co
mbined the peptide-functionalized hydrogel technology with bioinfor
matics to construct a library co
mposed of 12 different RGD peptides, including 6 unexplored RGD peptides, to develop culture substrates for hiPSC-derived cardio
myocytes (hiPSC-CMs), a cell type known for poor adhesion to synthetic substrates. 2 out of 6 unexplored RGD peptides showed substantial activities to support hiPSC-CMs. A
mong the
m, PMQKM
m>RGDm>VFSP fro
m la
minin β4 subunit was found to support the highest adhesion and sarco
mere for
mation of hiPSC-CMs. With bioinfor
matics, the peptide-functionalized hydrogel
microarrays accelerate the discovery of novel biological ligands to develop bio
materials for ste
m cell and tissue engineering applications.
Statement of Significance
In this manuscript, we described the development of a robust approach to prepare peptide-functionalized synthetic hydrogel microarrays. Combined with bioinformatics, this technology enables us to rapidly identify novel biological ligands for the development of the next generation of functional biomaterials for stem cell and tissue engineering applications.