A photolithographic method to create cellular micropatterns.
Here we describe a simple and rapid system for creation of patterned cell culture substrates. This technique is based on (1) printing a mask on a standard overhead transparency, (2) coating a thin layer of a photocrosslinkable chitosan on a slide, (3) exposing the slide and mask to ultraviolet (UV) light, and (4) rinsing the uncrosslinked polymer to expose the underlying cell-repellent patterns. Photocross-linkable chitosan does not require photoinitiators, it is non-toxic and forms flexible, biocompatible hydrogel upon short ( approximately min) UV exposure. Patterns of various shapes (lanes, squares, triangles, circles) were created on two surfaces commonly used for cell culture: glass and tissue culture polystyrene. The pattern size could be varied with a mum resolution using a single mask and varying UV exposure time. Cardiac fibroblasts formed stable patterns for up to 18 days in culture. Cardiomyocytes, patterned in lanes 68-99 microm wide, exhibited expression of cardiac Troponin I, well developed contractile apparatus and they contracted synchronously in response to electrical field stimulation. Osteoblasts (SAOS-2) localized in the exposed glass regions (squares, triangles, or circles; 0.063-0.5mm(2)). They proliferated to confluence in 5 days, expressed alkaline phosphatase and produced a mineralized matrix.
Karp
Jeffrey M
JM
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, 02139, USA.
Yeo
Yoon
Y
Geng
Wenlinag
W
Cannizarro
Christopher
C
Yan
Kenny
K
Kohane
Daniel S
DS
Vunjak-Novakovic
Gordana
G
Langer
Robert S
RS
Radisic
Milica
M
D000818:Animals; D000831:Animals, Newborn; D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000379:methods*; D016764:Cell Polarity; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D006801:Humans; D008422:Materials Testing; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology; D010006:Osteoblasts / Q000166:cytology* / Q000502:physiology; D010781:Photography / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*
D001672:Biocompatible Materials; D048271:Chitosan