Retinal pigment epithelial cell behavior is modulated by alterations in focal cell-substrate contacts.
OBJECTIVE
To investigate how the cellular behavior of cultured retinal pigment epithelial (RPE) cells was affected by the manipulation of early focal contact.


METHODS
To manipulate early focal contact, a reduced focal cell-substrate contact area on the micropatterned surfaces was implemented by microfabrication with polydimethylsiloxane (PDMS). The micropatterned PDMS surfaces had a circular pillar with a diameter of 5 microm. The human retinal pigment epithelial cell line, ARPE-19, was seeded onto the fibronectin-coated PDMS surfaces. Cell adhesion, growth, cell cycle, morphology, and interleukin-6 (IL-6) expression were observed for 3 weeks.


RESULTS
The fluorescent images of localized vinculin and actin stress fibers appeared to be more prominent on smooth PDMS surfaces. Although there was no significant effect on cell adhesion, a statistically significant inhibition of cell cycle progression was observed for micropatterned PDMS surfaces. Similarly, micropatterned surfaces showed significantly less cell growth than that of smooth surfaces. Cultures over a period of 3 weeks showed a distinct cell-cell phenotype discrepancy. Furthermore, IL-6 mRNA and secreted protein induced by IL-1beta in ARPE-19 were downregulated on micropatterned PDMS surfaces.


CONCLUSIONS
Disturbed focal contact in ARPE-19 cells grown on micropatterned surfaces altered cell cycle, growth, morphology, and the expression of IL-6 in vitro.
Lim
Jung-Min
JM
School of Electrical Engineering and Computer Science, Seoul National University, Korea.
Byun
Sangwon
S
Chung
Seok
S
Park
Tai Hyun
TH
Seo
Jong-Mo
JM
Joo
Choun-Ki
CK
Chung
Hum
H
Cho
Dong-Il
DI
D000199:Actins / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D002453:Cell Cycle / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D004129:Dimethylpolysiloxanes; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins; D006801:Humans; D007150:Immunohistochemistry; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins; D010857:Pigment Epithelium of Eye / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012828:Silicones; D016596:Vinculin / Q000378:metabolism
D000199:Actins; D004129:Dimethylpolysiloxanes; D005353:Fibronectins; D015850:Interleukin-6; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D012828:Silicones; C048022:substrate adhesion molecules; D016596:Vinculin; C013830:baysilon