Gwon-Soo Jung, Jin-Kyu Park, Kyeong-Min Lee, Mi-Ae Kwon and Won-Bae Jeon
Posters: J Cell Sci Ther
Elastin-like polypeptides (ELPs) are consist of VPGXG pentapeptide (X is the guest position for any amino acid except proline) derived from the VPGVG sequence found in a natural matrix protein tropoelastin. Neuronal cells require contact sites within their surrounding matrix for not only initial cell attachment, but long-term diff erentiation and neuronal morphogenesis as well. In natural ECM environment, the cell adhesions sequences or domains found in various types of ECM proteins provide contact sites and regulate neuronal cell behaviors through the interaction with cell-surface receptors. Among them, Arg-Gly-Asp (RGD) tripeptide derived from fi bronectin, vitronectin and collagen is a potent integrin binding ligand associated with adhesionmediated cell motility and neurite elongation during neuronal cell diff erentiation. In this study, an RGD-containing elastin-mimetic polypeptide TGPG [VGRGD (VGVPG) 6] 20WPC is referred to as [RGD-V6] and protein surfaces were prepared by isothermal adsorption of [RGDV6]20 and fi bronectin on tissue culture polystyrene. Th e eff ects of protein coated surfaces at 4ºC on N2a cellular behaviors were quantitatively measured to investigate its potential feasibility as a biomimetic analogue of natural matrix proteins. Th e [RGD-V6]20 polypeptide enhanced N2a cell attachment, and consistent with the cell adhering activity, increased migration speed and neurite outgrowth in a manner similar to native fi bronectin. Also, the expression levels of α3, α5, αv, β1, and 3 integrin subunits in the N2a cells grown on [RGDV6]20 were quantifi ed by RT-PCR, and were statistically not diff erent from those cultured on polystyrene surface. Adhesion-mediated enhancement in cell migration and diff erentiation is generally desired in tissue regeneration. In this context, the [RGD-V6]20 polypeptide can provide a bioactive matrix for neuronal cell engineering or to improve long- term biocompatibility of tissue implant devices. But, the eff ects of protein coated surfaces at 37ºC on N2a cellular behaviors are diff erent compared with those of protein coated surfaces at 4ºC. Cells form the spheroid and are not diff erentiated. Th ese changes are alike with results of human neuroblastoma, SH-SY5Y culture on the protein coated surfaces at 37ºC. Morphology of SH-SY5Y is similar with that of N2a. Moreover, the expression levels of integrin subunits also are decreased on the protein coated surfaces at 37ºC. Th refore, we concluded RGD-functionalized ELP aff ect diff erently morphology and diff erentiation of neuroblastoma cell according to protein coating temperature via the expression of integrins. We think that the diff erences of cellular morphology according to the RGD-functionalized ELP coating temperature are used as experimental tool in other experiments.
Dr. Jung completed his Ph.D at the age of 28 years from Keimyung University, College of Medicine in South Korea. She has published 4 papers. Now, she works at Daegu Gyeongbuk Institute of Science and Technology as a postdoctoral researcher.
