TY - JOUR
T1 - Covalent attachment of an Arg-Gly-Asp sequece peptide to derivatizable polyacrylamide surfaces
T2 - Support of fibroblast adhesion and long-term growth
AU - Brandley, Brian K.
AU - Schnaar, Ronald L.
N1 - Funding Information:
’ This work was supported by National Institutes of Health Grants HD 14010 and HD 20527 to R.L.S. and by Biomedical Research Support Grant RR5378 to B.K.B. R.L.S. (to whom correspondence should be addressed) is the recipient of an American Cancer Society Faculty Research Award (FRA-280). 2 Abbreviations used: BSA, bovine serum albumin; DMEM, Dulbecco’s modified Eagle medium; Hepes, 4-(2-hydroxyethyl)-I-piperazineethanesulfonic acid; IgG, immunoglobulin G; LDH, lactate dehydrogenase; PBS, phosphate-buffered saline; RGD, arginine-gly-tine-aspartic acid,N-6, N-succinimidyels tero f acryla- midohexanoic acid.
PY - 1988/7
Y1 - 1988/7
N2 - A synthetic nonapeptide (Tyr-Ala-Val-Thr-Gly-Arg-Gly-Asp-Ser), which includes the adhesive Arg-Gly-Asp (RGD) sequence, was covalently immobilized on chemically well-defined polyacrylamide gel surfaces utilizing N-succinimidyl active esters. The amount of peptide immobilized varied linearly with the concentration added to the gels. Immobilization was ∼80% efficient (based on peptide added), resulting in up to 17.5 nmol peptide/cm2 gel surface. Balb/c 3T3 mouse fibroblast cells adhered readily to peptide-derivatized surfaces, even in the absence of serum. Furthermore, surfaces derivatized with 2 nmol peptide/cm2 gel supported long-term fibroblast growth at a rate and to an extent comparable to that on tissue culture plastic. Surfaces derivatized with a control nonapeptide having no RGD sequence were nonsupportive of cell attachment or growth. The immobilization technology used to derivatize the gel surfaces with adhesive nonapeptide can be modified to allow coderivatization with proteins, glycoproteins, glycosides, or other amine-containing compounds to test their effects on long-term cell behaviors.
AB - A synthetic nonapeptide (Tyr-Ala-Val-Thr-Gly-Arg-Gly-Asp-Ser), which includes the adhesive Arg-Gly-Asp (RGD) sequence, was covalently immobilized on chemically well-defined polyacrylamide gel surfaces utilizing N-succinimidyl active esters. The amount of peptide immobilized varied linearly with the concentration added to the gels. Immobilization was ∼80% efficient (based on peptide added), resulting in up to 17.5 nmol peptide/cm2 gel surface. Balb/c 3T3 mouse fibroblast cells adhered readily to peptide-derivatized surfaces, even in the absence of serum. Furthermore, surfaces derivatized with 2 nmol peptide/cm2 gel supported long-term fibroblast growth at a rate and to an extent comparable to that on tissue culture plastic. Surfaces derivatized with a control nonapeptide having no RGD sequence were nonsupportive of cell attachment or growth. The immobilization technology used to derivatize the gel surfaces with adhesive nonapeptide can be modified to allow coderivatization with proteins, glycoproteins, glycosides, or other amine-containing compounds to test their effects on long-term cell behaviors.
KW - cell adhesion
KW - cell culture techniques
KW - fibronectin
KW - immobilized peptides
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U2 - 10.1016/0003-2697(88)90442-3
DO - 10.1016/0003-2697(88)90442-3
M3 - Article
C2 - 3189771
AN - SCOPUS:0023687433
SN - 0003-2697
VL - 172
SP - 270
EP - 278
JO - Analytical biochemistry
JF - Analytical biochemistry
IS - 1
ER -