A concept for miniaturized 3-D cell culture using an extracellular matrix gel.
This paper presents a novel method to embed, anchor, and cultivate cells in a controlled 3-D flow-through microenvironment. This is realized using an etched silicon pillar flow chamber filled with extracellular matrix (ECM) gel mixed with cells. At 4 degrees C, while in liquid form, ECM gel is mixed with cells and injected into the chamber. Raising the temperature to 37 degrees C results in a gel, with cells embedded. The silicon pillars both stabilize and increase the surface to volume ratio of the gel. During polymerization the gel shrinks, thus creating channels, which enables perfusion through the chip. The pillars increase the mechanical stability of the gel permitting high surface flow rates without surface modifications. Within the structure cells were still viable and proliferating after 6 days of cultivation. Our method thus makes it possible to perform medium- to long-term cultivation of cells in a controlled 3-D environment. This concept opens possibilities to perform studies of cells in a more physiological environment compared to traditional 2-D cultures on flat substrates.
Frisk
Thomas
T
Microsystems Laboratory, School of Electrical Engineering, Royal Institute of Technology, Stockholm, Sweden. Thomas.Frisk@s3.kth.se
Rydholm
Susanna
S
Andersson
Helene
H
Stemme
Göran
G
Brismar
Hjalmar
H
D000818:Animals; D019556:COS Cells; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002470:Cell Survival; D002522:Chlorocebus aethiops; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000737:chemistry*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005782:Gels / Q000737:chemistry; D007797:Laminin / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000379:methods; D008904:Miniaturization / Q000379:methods*; D011509:Proteoglycans / Q000737:chemistry
D004338:Drug Combinations; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005782:Gels; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; C085925:calcein AM; D003094:Collagen