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Figure 44.4. Competitive assay for HUVEC detachment from RGDS-
immobilized temperature-responsive culture surfaces by soluble peptide
addition. Soluble RGDS (100 mM) was added to cells on RGDS-IC(X) after a
24-hcultureat37 Cwithoutserum.CIPAAmcompositionswere1(opencir-
cle),3(triangle),5(square),and10mol%(diamond).Asacomparison,cells
culturedonTCPSfor24hat37 Cwithserumweredetachedat20 C(closed
circle). Reprinted with permission from Ref. 40. Copyright 2004 American
Chemical Society.
Immobilized RGDS contributed to the enhancement of cell
spreading, likely mediated by cell integrin. In contrast, the con-
trolarginine-glycine-glutamicacid-serine(RGES)-immobilizedsur-
faces(RGES-IC(X))didnotdemonstratecellspreadingenhancement
against immobilized peptide density because they have no inte-
grin biospecificity as a result of the substitution of glutamic acid
(E) for aspartic acid (D). Figure 44.4 shows spread human vascu-
lar endothelial cell (HUVEC) numbers on various substrates after 6-
hour and 24-hour cultures with or without serum at 37 C. Although
the addition of dilute serum facilitates HUVEC spreading on TCPS,
the RGDS modified poly(IPAAm-co-CIPAAm) surface dramatically
promoted cell spreading under serum-free conditions. In contrast,
little spreading improvement was observed on the RGDS-modified
poly(IPAAm-co-AAc) surface on which peptides were immobilized
via another co-monomer, AAc, in place of CIPAAm. As shown in Fig.
44.4, significant differences in HUVEC spreading on poly(IPAAm- co -
CIPAAm)-grafted surfaces (CIPAAm; 1 mol % in feed) before and
after RGDS immobilization after a 6-hour culture without serum at
37 C are readily evident.
 
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