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. 2011 Mar 30;5(1):13402.
doi: 10.1063/1.3516657.

Cell-enclosing gelatin-based microcapsule production for tissue engineering using a microfluidic flow-focusing system

Shinji Sakai, Sho Ito, Hitomi Inagaki, Keisuke Hirose, Tomohiro Matsuyama, Masahito Taya, Koei Kawakami

Cell-enclosing gelatin-based microcapsule production for tissue engineering using a microfluidic flow-focusing system

Shinji Sakai et al. Biomicrofluidics. .

Abstract

Gelatin-based microcapsule production using a microfluidic system and the feasibility of the resultant microcapsules for constructing spherical tissues surrounded by heterogeneous cells were studied. The first cell-encapsulation and subsequent cell-enclosing microparticle encapsulation were achieved using a microfluidic flow-focusing droplet production system. A hollow-core structure of about 150 μm in diameter was developed by incubating the resultant microparticles at 37 °C, which induced thermal melting of the enclosed unmodified gelatin microparticles. Mammalian cells filled the hollow-cores after 4 days of incubation. A cell layer on the cell-enclosing microcapsules was developed by simply suspending the microcapsules in medium containing adherent fibroblast cells. This method may prove useful for the generation of gelatin microcapsules using a microfluidic system for formation of artificial tissue constructs.

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Figures

Figure 1
Figure 1
Schematic illustration of a flow-focusing microdroplet production device.
Figure 2
Figure 2
Thermal gelation and melting temperatures of unmodified gelatin and Gelatin-Phs.
Figure 3
Figure 3
Effect of flow rate of liquid paraffin on diameter of 3% (w∕v) Gelatin-HPh droplets. Bars: mean±SD (n=50). Flow rate of Gelatin-HPh solution was fixed at 0.05 ml∕min.
Figure 4
Figure 4
Microphotographs of rat adipose-derived cells-enclosing ECGelatin-HPh microparticles (a) just after encapsulation of unmodified gelatin microparticles, after (b) 1 and (c) 4 days of incubation, and 1 day after seeding L929 cells on HepG2 cell-enclosing ECGelatin-HPh microcapsules at (d) low magnification and (e) high magnification. The arrow in panel (a) indicates the cell-enclosing unmodified gelatin microparticles without ECGelatin-HPh gel membrane. Bars indicate the scale: either 100 or 500 μm.
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