Directed cell growth on protein-functionalized hydrogel surfaces.
Biochemical surface modification has been used to direct cell attachment and growth on a biocompatible gel surface. Acrylamide-based hydrogels were photo-polymerized in the presence of an acroyl-streptavidin monomer to create planar, functionalized surfaces capable of binding biotin-labelled proteins. Soft protein lithography (microcontact printing) of proteins was used to transfer the biotinylated extracellular matrix proteins, fibronectin and laminin, and the laminin peptide biotin-IKVAV, onto modified surfaces. As a biological assay, we plated LRM55 astroglioma and primary rat hippocampal neurons on patterned hydrogels. We found both cell types to selectively adhere to areas patterned with biotin-conjugated proteins. Fluorescence and bright-field modes of microscopy were used to assess cell attachment and cell morphology on modified surfaces. LRM55 cells were found to attach to protein-stamped regions of the hydrogel only. Neurons exhibited significant neurite extension after 72h in vitro, and remained viable on protein-stamped areas for more than 4 weeks. Patterned neurons developed functionally active synapses, as measured by uptake of the dye FM1-43FX. Results from this study suggest that hydrogel surfaces can be patterned with multiple proteins to direct cell growth and attachment.
Hynd
Matthew R
MR
Laboratory of Nervous System Disorders, Wadsworth Center, New York State Department of Health, Albany, NY 12201-0509, USA. mhynd@wadsworth.org
Frampton
John P
JP
Dowell-Mesfin
Natalie
N
Turner
James N
JN
Shain
William
W
D000818:Animals; D001672:Biocompatible Materials; D019921:Biotinylation; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000502:physiology*; D005353:Fibronectins; D020100:Hydrogels; D007700:Kinetics; D007797:Laminin; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D011506:Proteins / Q000502:physiology*; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D051381:Rats; D013499:Surface Properties
D001672:Biocompatible Materials; C073804:FM1 43; D005353:Fibronectins; D020100:Hydrogels; D007797:Laminin; D011506:Proteins; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds