Enhanced survival of neural cells embedded in hydrogels composed of collagen and laminin-derived cell adhesive peptide.
To develop biomaterials that serve to improve the survival of neural cells transplanted into central nervous tissues, type I collagen-based hydrogels were prepared as a cell carrier. The hydrogels were modified with a laminin-derived peptide that is known to have an affinity for alpha3beta1 integrin, to transduce antiapoptotic signaling in embedded cells. For the modification of collagen, the peptide was fused to the N- or C-terminus, or both termini of a collagen-binding polypeptide domain by means of recombinant DNA technology. The chimeric proteins were characterized by polyacrylamide gel electrophoresis and circular dichroism spectroscopy, while binding of chimeric proteins to collagen-coated substrates was verified by surface plasmon resonance analysis under physiological conditions. Cell culture assays revealed that the adhesion of neurosphere-forming cells to collagen-coated polystyrene surfaces was significantly promoted by the incorporation of the chimeric proteins in a peptide-density dependent manner. The live/dead assays for cells cultured for 24 or 48 h in the hydrogels revealed that peptide incorporation improved the survival of cells embedded in collagen hydrogels. These results suggest that collagen hydrogel containing the laminin-derived peptide provides microenvironments suitable for the survival of neural cells.
Institute for Frontier Medical Sciences, Kyoto University, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Makiko
M
Hiraoka
Koichi
K
Kato
Tadashi
T
Nakaji-Hirabayashi
Hiroo
H
Iwata
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000166:cytology* / Q000187:drug effects*; D002448:Cell Adhesion / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen / Q000378:metabolism*; D020100:Hydrogels; D007797:Laminin / Q000737:chemistry*; D008958:Models, Molecular; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry* / Q000235:genetics; D011137:Polystyrenes / Q000737:chemistry; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects; D013499:Surface Properties; D014841:von Willebrand Factor / Q000737:chemistry / Q000378:metabolism
D020100:Hydrogels; D007797:Laminin; D009842:Oligopeptides; D011137:Polystyrenes; D011993:Recombinant Fusion Proteins; D014841:von Willebrand Factor; D003094:Collagen