Insulinoma cell line (MIN6) adhesion and spreading mediated by Arg-Gly-Asp (RGD) sequence conjugated in thermo-reversible gel.
We have functionalized gels with a putative cell-binding (-Arg-Gly-Asp-) (RGD) domain in an effort to regulate mammalian cell behavior in cells entrapped with gel. Adhesion molecules composed of Gly-Arg-Gly-Asp-Ser (GRGDS) peptides and cell recognition ligands were inculcated into thermo-reversible hydrogel composed of N-isopropylacrylamide, with a small amount of succinyl poly(ethylene glycol) (PEG) acrylate (MW 2000) used as a biomimetic extracellular matrix (ECM). The GRGDS-containing p(NiPAAm-co-PEG) copolymer gel was studied in vitro for its ability to promote cell spreading and to increase the viability of cells by introducing PEG spacers. Hydrogel lacking the adhesion molecules proved to be a poor ECM for adhesion, permitting only a 20% spread of the seeded cells after 10 d. When PEG spacer arms, immobilized by a peptide linkage, had been integrated into the hydrogel, conjugation of RGD promoted cell spread by 300% in a 28-d trial. In addition, in a serum-free medium, only GRGDS peptides conjugated with the spacer arm were able to promote cell spread.
Park
Keun-Hong
KH
College of Medicine, Pochon CHA University, Cell and Gene Therapy Research Institute, 605 Yeoksam 1-dong, Kangnam-gu, Seoul 135-081, Korea.
Na
Kun
K
Jung
Seung Yong
SY
Kim
Sung Won
SW
Park
Kyu Hyung
KH
Cha
Kwang Yul
KY
Chung
Hyung-Min
HM
D000327:Adsorption; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry*; D007340:Insulinoma / Q000473:pathology / Q000503:physiopathology; D007515:Islets of Langerhans / Q000166:cytology* / Q000502:physiology*; D008422:Materials Testing; D051379:Mice; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D023822:Tissue Engineering / Q000379:methods*
D020099:Coated Materials, Biocompatible; D020100:Hydrogels; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid