Fabrication of cellular spheroids is critical for creating functional tissue units and investigating the mechanism of tumorigenesis, development, and intercellular and cell–matrix interactions in vitro. Herein, we developed a novel, simple, and facile method for cell spheroid fabrication by using polyacrylamide/gelatin methacrylate (PA/GelMA) hydrogel composites. Arrays of Michigan Cancer Foundation-7 (MCF-7) breast cancer cell spheroids can be easily formed by tuning the GelMA composition. The shape and size of cell spheroids can be also well controlled by regulating cell seeding density and culturing time. All these results suggested that this simple and facile platform can serve as a useful tool to generate three-dimensional (3D) cell spheroids and can be integrated within high-throughput drug screening platforms, which will be of great help in engineering functional tissue models and regenerative medicines.

References

1.
Tekin
,
H.
,
Anaya
,
M.
,
Brigham
,
M. D.
,
Nauman
,
C.
,
Langer
,
R.
, and
Khademhosseini
,
A.
,
2010
, “
Stimuli-Responsive Microwells for Formation and Retrieval of Cell Aggregates
,”
Lab Chip
,
10
(
18
), pp.
2411
2418
.
2.
Tong
,
J. Z.
,
Bernard
,
O.
, and
Alvarez
,
F.
,
1990
, “
Long-Term Culture of Rat Liver Cell Spheroids in Hormonally Defined Media
,”
Exp. Cell Res.
,
189
(
1
), pp.
87
92
.
3.
Hober
,
C.
,
Benhamou
,
P. Y.
,
Watt
,
P. C.
,
Watanabe
,
Y.
,
Nomura
,
Y.
,
Stein
,
E.
,
Brunicardi
,
F. C.
, and
Mullen
,
Y.
,
1997
, “
A New Culture Method for Human Pancreatic Islets Using a Biopore Membrane Insert
,”
Pancreas
,
14
(
2
), pp.
199
204
.
4.
Sutherland
,
R. M.
,
1988
, “
Cell and Environment Interactions in Tumor Microregions: The Multicell Spheroid Model
,”
Science
,
240
(
4849
), pp.
177
184
.
5.
de Ridder
,
L.
,
Cornelissen
,
M.
, and
de Ridder
,
D.
,
2000
, “
Autologous Spheroid Culture: A Screening Tool for Human Brain Tumour Invasion
,”
Crit. Rev. Oncol. Hemat.
,
36
(
23
), pp.
107
122
.
6.
Mignatti
,
P.
,
Robbins
,
E.
, and
Rifkin
,
D. B.
,
1986
, “
Tumor Invasion Through the Human Amniotic Membrane: Requirement for a Proteinase Cascade
,”
Cell
,
47
(
4
), pp.
487
498
.
7.
Mohr
,
J. C.
,
de Pablo
,
J. J.
, and
Palecek
,
S. P.
,
2006
, “
3-D Microwell Culture of Human Embryonic Stem Cells
,”
Biomaterials
,
27
(
36
), pp.
6032
6042
.
8.
Hwang
,
Y.-S.
,
Chung
,
B. G.
,
Ortmann
,
D.
,
Hattori
,
N.
,
Moeller
,
H.-C.
, and
Khademhosseini
,
A.
,
2009
, “
Microwell-Mediated Control of Embryoid Body Size Regulates Embryonic Stem Cell Fate Via Differential Expression of WNT5a and WNT11
,”
Proc. Natl. Acad. Sci. USA
,
106
(
40
), pp.
16978
16983
.
9.
Tung
,
Y.-C.
,
Hsiao
,
A. Y.
,
Allen
,
S. G.
,
Torisawa
,
Y.-S.
,
Ho
,
M.
, and
Takayama
,
S.
,
2011
, “
High-Throughput 3D Spheroid Culture and Drug Testing Using a 384 Hanging Drop Array
,”
Analyst
,
136
(
3
), pp.
473
478
.
10.
Li
,
Y.
,
Huang
,
G.
,
Zhang
,
X.
,
Li
,
B.
,
Chen
,
Y.
,
Lu
,
T.
,
Lu
,
T. J.
, and
Xu
,
F.
,
2013
, “
Magnetic Hydrogels and Their Potential Biomedical Applications
,”
Adv. Funct. Mater.
,
23
(
6
), pp.
660
672
.
11.
Ling
,
K.
,
Huang
,
G.
,
Liu
,
J.
,
Zhang
,
X.
,
Ma
,
Y.
,
Lu
,
T. J.
, and
Xu
,
F.
,
2015
, “
Bioprinting-Based High-Throughput Fabrication of Three-Dimensional MCF-7 Human Breast Cancer Cellular Spheroids
,”
Engineering
,
1
(
2
), pp.
269
274
.
12.
EunáKim
,
Y.
,
SunáLee
,
K.
,
HyunáLee
,
S.
,
WookáOh
,
K.
, and
YoonáKang
,
J.
,
2011
, “
Generation of Core–Shell Microcapsules With Three-Dimensional Focusing Device for Efficient Formation of Cell Spheroid
,”
Lab Chip
,
11
(
2
), pp.
246
252
.
13.
Wu
,
L. Y.
,
Di Carlo
,
D.
, and
Lee
,
L. P.
,
2008
, “
Microfluidic Self-Assembly of Tumor Spheroids for Anticancer Drug Discovery
,”
Biomed. Microdevices
,
10
(
2
), pp.
197
202
.
14.
Dean
,
D. M.
,
Napolitano
,
A. P.
,
Youssef
,
J.
, and
Morgan
,
J. R.
,
2007
, “
Rods, Tori, and Honeycombs: The Directed Self-Assembly of Microtissues With Prescribed Microscale Geometries
,”
FASEB J.
,
21
(
14
), pp.
4005
4012
.
15.
Karp
,
J. M.
,
Yeh
,
J.
,
Eng
,
G.
,
Fukuda
,
J.
,
Blumling
,
J.
,
Suh
,
K.-Y.
,
Cheng
,
J.
,
Mahdavi
,
A.
,
Borenstein
,
J.
,
Langer
,
R.
, and
Khademhosseini
,
A.
,
2007
, “
Controlling Size, Shape and Homogeneity of Embryoid Bodies Using Poly (Ethylene Glycol) Microwells
,”
Lab Chip
,
7
(
6
), pp.
786
794
.
16.
Fukuda
,
J.
,
Khademhosseini
,
A.
,
Yeo
,
Y.
,
Yang
,
X.
,
Yeh
,
J.
,
Eng
,
G.
,
Blumling
,
J.
,
Wang
,
C. F.
,
Kohane
,
D. S.
, and
Langer
,
R.
,
2006
, “
Micromolding of Photocrosslinkable Chitosan Hydrogel for Spheroid Microarray and Co-Cultures
,”
Biomaterials
,
27
(
30
), pp.
5259
5267
.
17.
Moeller
,
H.-C.
,
Mian
,
M. K.
,
Shrivastava
,
S.
,
Chung
,
B. G.
, and
Khademhosseini
,
A.
,
2008
, “
A Microwell Array System for Stem Cell Culture
,”
Biomaterials
,
29
(
6
), pp.
752
763
.
18.
Cordey
,
M.
,
Limacher
,
M.
,
Kobel
,
S.
,
Taylor
,
V.
, and
Lutolf
,
M. P.
,
2008
, “
Enhancing the Reliability and Throughput of Neurosphere Culture on Hydrogel Microwell Arrays
,”
Stem Cells
,
26
(
10
), pp.
2586
2594
.
19.
Wood
,
D. K.
,
Weingeist
,
D. M.
,
Bhatia
,
S. N.
, and
Engelward
,
B. P.
,
2010
, “
Single Cell Trapping and DNA Damage Analysis Using Microwell Arrays
,”
Proc. Natl. Acad. Sci. USA
,
107
(
22
), pp.
10008
10013
.
20.
Zhang
,
A. P.
,
Qu
,
X.
,
Soman
,
P.
,
Hribar
,
K. C.
,
Lee
,
J. W.
,
Chen
,
S.
, and
He
,
S.
,
2012
, “
Rapid Fabrication of Complex 3D Extracellular Microenvironments by Dynamic Optical Projection Stereolithography
,”
Adv. Mater.
,
24
(
31
), pp.
4266
4270
.
21.
Li
,
Y.
,
Poon
,
C. T.
,
Li
,
M.
,
Lu
,
T. J.
,
Pingguan-Murphy
,
B.
, and
Xu
,
F.
,
2015
, “
Chinese‐Noodle‐Inspired Muscle Myofiber Fabrication
,”
Adv. Funct. Mater.
,
25
(
37
), pp.
5999
6008
.
22.
Tang
,
X.
,
Ali
,
M. Y.
, and
Saif
,
M. T. A.
,
2012
, “
A Novel Technique for Micro-Patterning Proteins and Cells on Polyacrylamide Gels
,”
Soft Matter
,
8
(
27
), pp.
7197
7206
.
23.
Nakayama
,
A.
,
Kakugo
,
A.
,
Gong
,
J. P.
,
Osada
,
Y.
,
Takai
,
M.
,
Erata
,
T.
, and
Kawano
,
S.
,
2004
, “
High Mechanical Strength Double‐Network Hydrogel With Bacterial Cellulose
,”
Adv. Funct. Mater.
,
14
(
11
), pp.
1124
1128
.
You do not currently have access to this content.