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Micro‐stamped ECM proteins enhance endothelial cell adhesion and directed growth
Author(s) -
Rubenstein David Alan,
Frame Mary D.
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1212-a
Subject(s) - fibronectin , extracellular matrix , laminin , chemistry , matrigel , adhesion , cell adhesion , matrix (chemical analysis) , polydimethylsiloxane , biophysics , microbiology and biotechnology , cell , materials science , nanotechnology , biology , biochemistry , chromatography , organic chemistry
Our goal was to evaluate microvascular endothelial cell growth within micro‐stamped patterns of extracellular matrix proteins (ECM). A combination of photo‐ and soft‐lithography was used. Features ~100μm deep and 50–150μm wide were fabricated from SU‐8 2100 photoresist. Polydimethylsiloxane imprints of features produced positive molds used to stamp extracellular matrix proteins (collagen I, IV, or laminin each at 5μg/mm2; fibronectin at 3.4μg/mm2) onto cleaned glass coverslips (n = 4 each). ECM was allowed to dry; the average stamp width was ~160μm, measured prior to cell seeding. Human dermal microvascular endothelial cells (ECs) were seeded at an initial density of 800 cells/cm2, and cultured for three days. We hypothesized that by day 3, ECs would have higher viability and density on the patterned matrices vs. on bare glass. On day 3, there were clearly visible bands of cells growing throughout the patterned matrices (~150–300μm wide). Cells were stained with calcein/ethidum and imaged to obtain viability and density on either the untreated bare glass, or along the stamped patterns. Cell viability was high (>90%) for all conditions independent of location, and was >95% along collagen I and fibronectin stamped patterns. Cell density increased by 4‐fold on glass (3,472±1,179 cells/cm2, mean±SD) and up to 16‐fold on the micro‐stamped proteins (fibronectin, 17,545±7,892; collagen IV, 15,381±7,031; collagen I, 12,498±8,583; laminin, 9,511±4,333 cells/cm2). Thus, concentrated growth of ECs was markedly enhanced within micro‐stamped ECM patterns suggesting either enhanced adhesion or proliferation. (NIH HL55492)