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DOI: 10.1002/adma.200700433

Monodisperse Alginate Hydrogel Microbeads for Cell

Encapsulation**
By Wei-Heong Tan and Shoji Takeuchi*
This Communication describes the production of monodisperse alginate hydrogel microbeads (94150 lm) using a
method that combines internal gelation method with T-junction droplet formation in microfluidic devices. The use of calcium carbonate (CaCO3) nano-particles allows internal gelation method to be applied to micro-scale production for the
first time, and microfluidic devices allow us to produce
microbeads with narrow size distributions. Our approach not
only allows easy control over bead size by varying flow
parameters, but also allows better monodispersity and control
over the shape of the hydrogel beads compared to conventional external gelation methods performed in microfluidic
devices. Both blank and cell encapsulating alginate hydrogel
beads of various shapes were successfully produced using this
approach in non-silanized/silanized poly(dimethylsiloxane)
(PDMS) devices. Also, we demonstrated that the gelation
conditions in our approach were mild enough to encapsulate
mammalian cells (Jurkat) without loss of their viability, and
studied the effect on cell viability with varying concentrations
of CaCO3.
Alginates are anionic polysaccharides extracted from seaweeds composing of b-D-mannuronic acid and a-L-guluronic
acid, and they form hydrogels with multivalent cations such as
Ca2+, Ba2+, or Fe3+. Due to their resemblance to the natural
extracellular matrix, alginate hydrogels have been employed
successfully in three-dimensional cell-hydrogel scaffolds for
tissue engineering,[1,2] and in the encapsulation of transplanted (allogenic or xenogeneic) cells in alginate hydrogel
beads. The alginate hydrogel provides an immunoisolation
barrier for the cells,[36] potentially allowing transplantation
without the need for immunosuppression. However, there
remain several issues that limit the use of cell encapsulation
technology for transplantation, namely: (i) Large bead size.
Size of these beads should approach that of the cells/tissues to

[*] Dr. S. Takeuchi, W.-H. Tan

CIRMM/IIS, The University of Tokyo, Institute of Industrial Science
4-6-1 Komaba, Meguro-ku, Tokyo 153-8505 (Japan)
E-mail: takeuchi@iis.u-tokyo.ac.jp
S. Takeuchi
PRESTO, JST
4-1-8 Honcho, Kawaguchi, Saitama 332-0012 (Japan)
[**] This work was supported by CREST from Japan Science and
Technology Agency and by Grants-in-Aid for Scientific Research on
Priority Areas (innovative nanoscience) from Ministry of Education,
Culture, Sports, Science, and Technology Japan. The authors also
acknowledge NanoMaterials Technology Pte Ltd. (Singapore) for
supplying the calcium carbonate nano-particles.

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be encapsulated to allow a high degree of permeability for

rapid exchange of nutrients into and waste out of the beads by
diffusion, thereby improving viability of cells/tissues.[4] However, conventional techniques[7] can only produce beads in the
order of several hundred micrometers; (ii) Wide size distribution. Monodisperse beads are important to accurately estimate the amount of transplanted cells introduced into a patient; (iii) Deformed beads. Morphology of the beads is also
an important parameter as deformed beads might reduce the
biocompatibility or make implantation more difficult since
beads carrying a tail are reported to cause fibrotic overgrowth
on surround tissue;[4,5] (iv) Process repeatability and reproducibility[6] in the production of uniform capsules have to be improved for actual clinical applications.
Highly reproducible monodisperse droplets can be obtained
using T-junction[8] or flow-focusing methods[9] in micro-devices, and several groups[10,11] have attempted to utilize such
techniques to produce alginate hydrogel beads; their strategies typically involved using micro-devices to form emulsions
of Na-alginate droplets in an organic phase, followed by the
external addition of Ca2+ ions in the form of calcium chloride
(CaCl2) solution. Rapid gelling behavior of alginate in this
external gelation approach made it difficult to produce welldefined and homogenous alginate hydrogel beads, as evident
from their images of the final alginate beads obtained.[10,11]
Moreover, the absence of surfactants in such systems often resulted in undesired fusion of Na-alginate droplets before gelation, resulting in high polydispersity of the beads.[10]
Here, we employ the internal gelation method[1214] to produce alginate hydrogel beads. This method involves dispersing
an insoluble (or slowly soluble) calcium complex in the Na-alginate solution. Upon pH reduction, Ca2+ ions are released from
the calcium complex, crosslinking the alginate to form a homogeneous hydrogel. Various groups[12,13] had tried but were unable to produce monodispersed alginate hydrogel beads with internal gelation using conventional emulsification methods. To
date, none had attempted to apply internal gelation in microdevices for the following reasons: (i) CaCO3 was often chosen
as the calcium complex for internal gelation. Commercial
CaCO3 powder (q = 2830 kg m3), consisting of grains with
f > 5 lm, will sediment 100 lm (typical microchannel dimensions) in water in a few seconds, rendering it an unsuitable technique for microfluidic devices; (ii) Large CaCO3 particle sizes
make it difficult to ensure a homogeneous dispersion within the
alginate, which is important for the production of small microbeads (f < 300 lm). In this work, we solved these problems
by using CaCO3 nano-particles (f 40 nm), drastically increasing

2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Adv. Mater. 2007, 19, 26962701

2H+ + CaCO3 Ca2+ + H2O + CO2

(1)

Ca2+ + 2Na+ Alg Ca2+ (Alg)2 + 2Na+

(2)

To extract these alginate hydrogel beads from the corn oil,

we first collected the emulsion in a microtube, and aspirated
off the excess oil after the beads had settled down. Hexane
was subsequently added to dissolve the residual corn oil and
at the same time quenching the gelation reaction, resulting in
the re-suspension of beads in hexane. Excess hexane was
again aspirated off before washing buffer was added. After
separating the beads into the washing buffer by centrifuga-

Figure 1. Schematic view of micro-device for production of alginate hydrogel microbeads. Droplets of Na-alginate containing CaCO3 nano-particles are formed at the T-junction. For cell encapsulation, cells are added
to the Na-alginate solution before introducing it into the device. Downstream of the T-junction, a stream of acidic oil merges with the main
stream. This pH reduction releases Ca2+, which reacts with Na-alginate
to form Ca-alginate gel downstream.

Adv. Mater. 2007, 19, 26962701

tion, we disposed off the supernatant and resuspended the

beads in cell culture medium. Centrifuging, decanting and resuspending in cell culture medium were repeated twice for
rinsing. This whole extraction process was accomplished within 15 min.
Figure 2a shows how the size or volume of the hydrogel
beads can be controlled by the flow rate of the continuous
c). At low CaCO3 and acetic acid concentrations, we
phase (Q
obtained spherical beads with diameters ranging from
94112 lm for the range of flow rates tested. These alginate
hydrogel beads exhibited a narrow size distribution with coefficient of variation (C.V.) < 2.9 %. C.V. is defined as the ratio
between the standard deviation and the mean. A C.V. of less
than 5 % is the commonly accepted definition of monodispersity.[9] As gelation is initiated from within the Na-alginate
droplets, the shape of these beads is very well preserved
(C.V.AR < 2.4 %, Fig. 2b and c) compared to external gelation
techniques[10,11] where addition of Ca2+ by fusion inherently
deforms the droplets. Theoretically, smaller beads can be produced at higher flow rates, but we experienced leakage with
c > 0.6 mL h1. We believe modificaour PDMS devices at Q
tions to the design (e.g., shortening the channel length) or
switching to devices that can withstand higher pressures (e.g.,
channels in a solid slab of PDMS[9] or silicon-glass based devices) should enable us to produce smaller hydrogel beads.
Xu et al.[15] reported solidifying droplets of monomer fluids
in microchannels by UV-photopolymerization to produce
beads of various shapes. They obtained nonspherical droplets
when ds is larger than at least one of the dimensions of the
outlet channel. ds refers to the diameter of an undeformed
droplet of volume, V, and is given by (6 V/p)1/3. The reason we
obtained spherical beads with diameters greater than the
height of the channel is because at these mild conditions, the
time needed for gelation is long. As a result, the droplets did
not gelate while under confinement in the microchannels; instead, these initially deformed droplets underwent shape relaxation into spheres before fully gelating into hydrogels outside of the micro-device. To produce non-spherical alginate
hydrogel beads with our system, the droplets must gelate sufficiently in their deformed state while confined in the microchannels. We can achieve this by manipulating 2 parameters
residence time (s) and reaction rate (v). s is determined by
the flow rate and the length of the channel (in our device,
length of the outlet channel is 32 mm); flow rate not only
affects s but also directly affects the droplets volume and
hence it is more convenient to control s by varying channel
length. However, as pressure difference increases proportionally to channel length, there is a limit to the maximum channel length that can be designed before leakage failure occurs.
On the other hand, v is determined by temperature and reactants concentration. As temperature control is difficult to implement, we will demonstrate the formation of discoid alginate hydrogel beads by increasing the concentration of the
reactants. By doubling the concentration of CaCO3 and increasing the acetic acid concentration by 4 times, we obtained
discoid beads with aspect ratio (AR) of 1.2 at 0.2 mL h1

2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

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the sedimentation time to the order of 104 s since the sedimentation velocity, U, is directly proportional to f2.
Figure 1 shows the design of the microfluidic device. In our
experiment, we dispersed droplets of Na-alginate in RPMI
(without NaHCO3) solution containing CaCO3 nano-particles
in a continuous phase of corn oil at a room temperature of
25 C. Syringe pumps were used to infuse fluids into the microfluidic device fabricated in PDMS using soft lithography
techniques. Droplets were generated at a T-junction, where
the narrow channel had a 50 48 lm2 cross section and the
main channel had a 150 48 lm2 cross section. Corn oil with
lecithin (2 % w/w) sheared off droplets of Na-alginate solution containing insoluble CaCO3 nano-particles one at a time
to generate an inverse emulsion with a narrow distribution in
droplet size. Lecithin was added to the corn oil to stabilize the
droplets against coalescence, thereby preserving the monodispersity of the droplets. Further downstream, acetic acid dissolved in corn oil (2 % w/w lecithin) was introduced, and this
acidic oil mixed with oil flowing in the main stream; the
acetic acid, which dissolved readily in both polar and non-polar solvents, then diffused into the aqueous Na-alginate droplets.[1214] This pH reduction (caused by protons diffusing into
the aqueous phase) released Ca2+ ions from the insoluble calcium complex (Eq. 1[14]), causing gelation (Eq. 2[14]).

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walls (Fig. 3a). However, when we attempted to encapsulate mammalian

cells (Jurkat) in alginate hydrogel beads
with the same system, we observed wetting of the PDMS channel (Fig. 3b) by
the Na-alginate solution. Initially, droplets were sheared off at the T-junction,
but as the front of the dispersed phase
gradually advanced downstream due to
wetting, droplets were sheared off
farther and farther downstream. Eventually, the stream of acidic oil caused
the front of the Na-alginate stream to
gelate and droplets were randomly
pinched off, producing polydisperse
droplets. This phenomenon was observed at all the flow rates tested. As
the main difference between producing
blank and cell encapsulating hydrogel
beads was the presence of cells, we postulate that this wetting of the channel
walls is due to the deposition of proteins, carbohydrates and cell debris,
which changes the surface energy of the
channel walls gradually from hydrophobic to hydrophilic. Silanizing the walls
of the microchannel solved this problem
(Fig. 3c). Figure 4a shows how the size
of the cell encapsulating hydrogel beads
c. Beads with length/diamvaries with Q
eter ranging from 100150 lm were obFigure 2. a) Plot of the size of alginate hydrogel microbeads versus flow rate of the continuous
phase (corn oil with lecithin (2 % w/w)) for different concentrations of reactants. The rate of flow
tained for the range of flow rates tested.
1
of the disperse phase, Qd, was 20 lL h . b) Plot of the AR of alginate hydrogel microbeads versus
All
the
beads
were
spherical
flow rate of the continuous phase for different concentrations of reactants. AR is defined as the ra(AR
<
1.03,
C.V.
<
3.0
%)
with
a narAR
tio of the length (l) to the width (w) of the bead. (see image (d)) Images (c)-(f) show the beads
row size distribution (C.V. < 3.2 %)
formed at various flow rates indicated in (a). These beads were extracted from corn oil and resuspended in cell culture medium. Inset in (d) shows 3 beads being stacked together, and the thickapart
for
those
produced
at
ness of each bead is 50 lm, which is close to the height of the channel. Scale bar applies to all im c = 0.2 mL h1, which were discoidal
Q
ages. g) Size distribution of the beads shown in (d). h) Size distribution of the beads shown in (f).
(AR = 1.2, C.V.AR = 5.9 %) with a wider
Both histograms were fitted with a Gaussian distribution.
size distribution (C.V. = 5.7 %). Although the dispersed phase did not wet
c = 0.2 mL h1, we observed from high
the channel walls at Q
(Fig. 2a, b, and d), where AR is defined as the ratio of the
speed camera images that the droplet shear-off point fluctulength (l) to the width (w) of the bead. These discoid beads
exhibit both narrow polydispersity in size (C.V. < 3.6 %,
ated between the corner of the T-junction, and at a point
Fig. 2a) and great uniformity in shape (C.V.AR = 2.65.1 %,
slightly downstream of the T-junction. The increase in polydis c = 0.2 mL h1 is probably due to the
persity of the beads at Q
Fig. 2b), which is not possible with conventional methods.

With increasing Qc, size of the beads gradually decreased and

fluctuation of the droplet shear-off point, although the reason
behind this fluctuation is not clear and the phenomenon is not
the beads adopted a more spherical shape (AR approaches
c. The procedure for extracting cell en c shortened s and offset the effect of highobserved at higher Q
unity); increase in Q
c. By
capsulating beads from corn oil was the same as that for blank
er v, as a result spherical beads were produced at high Q
beads except hexadecane was used instead of hexane. Hexane
tuning flow parameters and concentration of the reactants, we
is an excellent solvent for corn oil and has a low boiling point
can control both size and morphology of the alginate hydrogel
(69 C). Any residual hexane that had not been aspirated off
beads. These shape-controllable monodisperse hydrogel beads
would evaporate away, and thus less rinsing was required
can be used as carriers in drug delivery systems[16,17] when
when hexane was used. Nevertheless, when it was used to exenzymes and proteins are immobilized in the hydrogel.
tract cell encapsulating hydrogel beads, all the cells died. To
Production of blank alginate microbeads was performed in
maintain the viability of the cells, we had to replace hexane
PDMS devices without surface modifications to the channel

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2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Adv. Mater. 2007, 19, 26962701

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Figure 3. Optical microscopy images of droplet formation in the microfluidic device taken with high speed camera. Droplet formation of (a)
Na-alginate solution containing only CaCO3 for the production of blank
alginate hydrogel microbeads in a non-silanized PDMS device.
Qc = 20 mL h1 and Qd = 20 lL h1. b) Na-alginate solution containing
both CaCO3 and Jurkat cells for the production of cell encapsulating alginate hydrogel microbeads in a non-silanized PDMS device. Wetting of
the PDMS channel caused the front of the Na-alginate stream to gradually advance downstream. Eventually, the front of the stream gelated and
droplets were randomly sheared off. Qc = 20 mL h1 and Qd = 10 lL h1.
c) Na-alginate solution containing both CaCO3 and Jurkat cells for the
production of cell encapsulating alginate hydrogel microbeads in a silanized PDMS device. Qc = 60 mL h1 and Qd = 20 lL h1.

with hexadecane. Hexadecane (solubility = 9.0 108 g/100 g

water at 25 C[18]) is almost completely immiscible with water
when compared to hexane (solubility = 9.5 104 g/100 g at
25 C[19]), which may explain why cells remained viable when
hexadecane was used. We also studied how the concentration
of CaCO3 affected the viability of the cells (Fig. 4b). Percentage of cells that remained alive after the encapsulation process increased from 19.3 % to 74.3 % when the CaCO3 concentration was increased from 1.14 to 9.10 mg mL1 solution.
Here, trypan blue was used to differentiate live from dead
cells. Figure 4c shows the cell encapsulating alginate hydrogel
beads, and Figure 4d shows the close-up of alginate beads
containing live/dead Jurkat cells after trypan blue was added.
Cells are selective in the compounds that pass through the
membrane; trypan blue is not absorbed in a live cell, but it traverses the membrane in a dead cell. Hence, only dead cells
will exhibit a distinctive blue color. Higher loading of CaCO3
leads to higher crosslink densities, resulting in stiffer alginate
hydrogels[20] that may help protect encapsulated cells from
mechanical stresses during preparation. Increase in viability
of the cells with the increase in CaCO3 concentration is also
attributed to the dual role played by CaCO3. Besides releasing Ca2+ when the pH lowers, CO32 is also released from
CaCO3. CO32 acts as a base, regulating the pH inside the
beads. Beads prepared with a higher concentration of CaCO3
essentially have a larger reserve of CO32 to prevent it from
becoming overly acidic. Thus, we believe that both the increase in mechanical strength of the hydrogel and the milder
internal environment subsequently translate to higher viability of the encapsulated cells.

Adv. Mater. 2007, 19, 26962701

Figure 4. a) Plot of the size and AR of cell encapsulating alginate hydrogel microbeads versus flow rate of the continuous phase (corn oil with lecithin (2 % w/w)). The rate of flow of the disperse phase, Qd, was
20 lL h1. The acid concentration was 1 lL acetic acid/mL oil. b) Graph
showing how the concentration of CaCO3 affects the viability of the encapsulated cells. Beads were produced at Qc = 30 mL h1 and
Qd = 20 lL h1 for all the cases. In all cell cultures, a certain proportion of
the cells will be dead. Here, the percentage of live cells after process is
defined as (Nalive,after/Nalive,before) 100% where Nalive,before and Nalive,after
refers to the percentage of cells alive before the experiment and percentage of cells alive after the experiment, respectively. Optical microscopy
image of (c) cell encapsulating beads produced at Qc = 30 mL h1 and
Qd = 20 lL h1, and d) cell encapsulating beads when trypan blue was
added to test for cell viability. Dead cells exhibit a distinctive blue color.

The use of CaCO3 nano-particles enabled us to combine internal gelation method with T-junction droplet formation to
produce alginate hydrogel microbeads, and modifications in
channel geometry should allow beads with a wider range of
sizes to be produced. The strategy described in the present
work has four significant advantages: 1) it offers easy control
over morphology and size by tuning flow parameters or concentration of reactants, 2) it allows beads with a narrow size
distribution and high uniformity in morphology to be pro-

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duced, 3) it may allow large scale production with parallelization, and 4) the process is mild enough to encapsulate mammalian cells. For cell encapsulation, results suggest that the
concentration of CaCO3 directly affects the viability of encapsulated cells; increasing the concentration increases the percentage of viable cells. To date, successful encapsulation of
mammalian cells in monodisperse alginate hydrogel microbeads (f < 300 lm) has not been reported, and we envision the
extension of the strategy reported here to the production of
monodisperse microbeads with modified alginates such as
those with arginine (R), glycine (G), and aspartic acid (D)
(RGD)-adhesive ligands incorporated,[20,21] which would enable the encapsulation of adhesive cells in the near future.
Moreover, a reliable method to produce cell encapsulating alginate hydrogel beads may allow us to form cell-based bead
arrays by harnessing the technology developed to handle and
manipulate microbeads, such as the dynamic microarray platform.[22] Such cell-based bead array systems will facilitate
studies of pathological and physiological phenomena in cells.
With enormous potential for cell-based diagnostic applications, drug testing, toxicology studies, drug delivery systems,
tissue engineering and regenerative medicine applications, we
believe our strategy and protocol are not just trivial advances
in alginate hydrogel microparticle technology, but will serve
to further accelerate the use of alginate in many fields.

Experimental
Materials: Sodium alginate (80120 mPa
S), corn oil, lecithin from
soybean, hexadecane, hexane, calcium chloride, potassium chloride,
hydrochloric acid (5 mol L1) and sodium hydroxide solution
(5 mol L1) were purchased from Wako Pure Chemical Industries, Ltd
(Japan). Sodium chloride and Tween 20 were purchased from Kanto
Chemical Co., Inc. (Japan). Acetic acid was purchased from Nacalai
Tesque, Inc. (Japan). Calcium carbonate (CaCO3) (NPCC-Fresh slurry, 8.0 wt % Slurry, Lot No: 19092006-1) was kindly supplied by NanoMaterials Technology Pte Ltd. (Singapore). RPMI-1640 Medium
powder (R6504), RPMI-1640 Medium (R8758)(hereafter RPMI medium), fetal bovine serum (FBS), HEPES solution, and L-GlutaminePenicillin-Streptomycin Solution (G.P.S.) (G6784) were purchased
from SigmaAldrich (USA). Trypan blue was obtained from MP Biomedicals, Inc. (France). All chemicals and reagents were used as procured without further purification. Unless otherwise specified, all
water used in the experiments refers to ultra pure water having a specific resistance of 18 MX cm that is obtained from a Millipore system.
Jurkat, Clone E6-1(Acute T cell leukemia, human) was purchased
from Dainippon Sumitomo Pharma Co., Ltd. (Japan) and cultured at
37 C and 5 % CO2. Cell culture medium consists of RPMI medium
supplemented with 10 % FBS, 1.1 mL/100 mL medium of G.P.S. and
HEPES.
Solution Preparation: Na-alginate ( 2 % w/w) in RPMI (without
NaHCO3) solution was prepared as follows: 12 g Na-alginate was
added very slowly with stirring to 288 g water to give a 4 % w/w solution. Once a homogeneous solution was obtained, the solution was
autoclaved. RPMI-1640 Medium (2 concentration, without NaHCO3), hereafter RPMI without NaHCO3 (2), was prepared by dissolving 10.4 g of RPMI-1640 Medium powder in 500 mL of water with
stirring. pH of the solution was adjusted to 4.0 with HCl to completely
dissolve the powder. 1.4 g NaCl was then added to replace NaHCO3.
(NaHCO3 was not added as instructed in the suppliers protocol as it
acts as a pH buffer, which will interfere with the gelation process.)

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The pH was then adjusted to 7.1 with NaOH before adding 5.5 mL
G.P.S. and 5.5 mL HEPES to the solution. The solution was then sterilized by filtration using a membrane with a porosity of 0.22 lm.
Finally, equal volumes of Na-alginate (4 % w/w) and RPMI without
NaHCO3 (2) were mixed to obtain a final solution of Na-alginate
( 2 % w/w) in RPMI (without NaHCO3) solution.
Na-alginate in RPMI (without NaHCO3) solution containing
CaCO3 was prepared as follows: CaCO3 nano-particles were collected
from the slurry by centrifugation. The process of resuspending the
particles in RPMI without NaHCO3 (1), centrifuging, and discarding
the supernatant was repeated twice for rinsing. CaCO3 nano-particles
were then suspended in 0.5 mL RPMI without NaHCO3 (1) and
added to 5 mL Na-alginate ( 2 % w/w) in RPMI (without NaHCO3)
solution. We then thoroughly mixed the solution to ensure even distribution of the particles in the solution.
Na-alginate in RPMI (without NaHCO3) solution containing
CaCO3 and cells was prepared as follows: Similar to the CaCO3 nanoparticles, Jurkat cells were collected from the culture medium by centrifugation. After the supernatant was discarded, 2 mL of Na-alginate
in RPMI (without NaHCO3) solution containing CaCO3 was added
and the cells were resuspended by gentle pipetting. Finally, we obtained a solution of Na-alginate ( 1.8 % w/w) in RPMI (without
NaHCO3) containing CaCO3 and cells. The final solution had a cell
concentration of (6.86 + 0.50) 106 cells mL1.
Washing buffer was prepared as follows: CaCl2 solution consisting
of CaCl2 (162 mM), KCl (2.68 mM) and phenol red (0.04 mM) was prepared. Equal volumes of CaCl2 solution and RPMI-1640 Medium
were mixed, and Tween 20 (0.1 % w/w) was added to the final solution.
Device Fabrication and Silanization: We fabricated the device in
PDMS (Sylgard 184, Dow corning, USA) using soft lithography techniques. Our master molds consisted of SU-8 (MicroChem Corp.,
USA) positive relief features fabricated on 2-in silicon wafers. PDMS
layer was then cast from SU-8 master mold, and access holes for inlets
and outlet were punched on the PDMS slab. Another PDMS slab was
prepared by curing PDMS in a Petri dish. Surfaces of the two PDMS
slabs were treated with oxygen plasma to activate the surfaces for irreversible sealing before they were permanently bonded together. We
then baked the device on a hotplate for 50 min at 76 C to strengthen
the bonding. Devices were not used for another 48 hr to allow the
PDMS channel walls to regain its hydrophobicity. Silanization: The
following procedures were carried out inside a fume hood as it is hazardous to work with silane. We added 0.1 mL (Tridecafluoro-1,1,2,2Tetrahydrooctyl)Trichlorosilane (Gelest, Inc., USA) to 1.4 mL 3 M
Performance Fluid PF-5060 (3M, Japan) to obtain a dilute silane solution. After bonding and baking the device for 50 mins at 76 C, the
device was exposed to oxygen plasma again before a small amount of
dilute silane solution was introduced into the device. Finally, the
device was baked for 3 hr at 90 C. Silanized devices can be used
immediately after baking.
Received: February 19, 2007
Revised: April 16, 2007
Published online: August 22, 2007

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Adv. Mater. 2007, 19, 26962701

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