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Technical Brief: Engineering Cell Microenvironments Using Novel Hydrogels

Polyacrylamide/GelMA Hydrogel Templates for Breast Cancer Cell Spheroids Fabrication

[+] Author and Article Information
Yuhui Li

The Key Laboratory of Biomedical Information
Engineering of Ministry of Education,
School of Life Science and Technology,
Xi'an Jiaotong University,
Xi'an 710049, China;
Bioinspired Engineering and Biomechanics
Center (BEBC),
Xi'an Jiaotong University,
Xi'an 710049, China
e-mail: yuhuili1988@gmail.com

Xiang Li

The Key Laboratory of Biomedical Information
Engineering of Ministry of Education,
School of Life Science and Technology,
Xi'an Jiaotong University,
Xi'an 710049, China;
Bioinspired Engineering and Biomechanics
Center (BEBC),
Xi'an Jiaotong University,
Xi'an 710049, China
e-mail: lixiang199109@gmail.com

Lu Zheng

Key Laboratory of Microgravity (National
Microgravity Laboratory),
Center of Biomechanics and Bioengineering,
Institute of Mechanics,
Chinese Academy of Sciences,
Beijing 100190, China;
Beijing Key Laboratory of Engineered
Construction and Mechanobiology,
Institute of Mechanics,
Chinese Academy of Sciences,
Beijing 100190, China
e-mail: taswzzx@163.com

Lin Wang

The Key Laboratory of Biomedical Information
Engineering of Ministry of Education,
School of Life Science and Technology,
Xi'an Jiaotong University,
Xi'an 710049, China;
Bioinspired Engineering and Biomechanics
Center (BEBC),
Xi'an Jiaotong University,
Xi'an 710049, China
e-mail: wanglin0527@126.com

Xiaohui Zhang

The Key Laboratory of Biomedical Information
Engineering of Ministry of Education,
School of Life Science and Technology,
Xi'an Jiaotong University,
Xi'an 710049, China;
Bioinspired Engineering and Biomechanics
Center (BEBC),
Xi'an Jiaotong University,
Xi'an 710049, China
e-mail: xiaohuizhang@mail.xjtu.edu.cn

Feng Xu

Mem. ASME
The Key Laboratory of Biomedical Information
Engineering of Ministry of Education,
School of Life Science and Technology,
Xi'an Jiaotong University,
Xi'an 710049, China;
Bioinspired Engineering and Biomechanics
Center (BEBC),
Xi'an Jiaotong University,
Xi'an 710049, China
e-mail: fengxu@mail.xjtu.edu.cn

1Corresponding author.

Manuscript received October 8, 2015; final manuscript received October 15, 2015; published online March 8, 2016. Assoc. Editor: Abraham Quan Wang.

J. Nanotechnol. Eng. Med 6(3), 034501 (Mar 08, 2016) (5 pages) Paper No: NANO-15-1088; doi: 10.1115/1.4031898 History: Received October 08, 2015; Revised October 15, 2015; Accepted October 19, 2015

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.

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References

Figures

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Fig. 1

Schematics of PA/GelMA hydrogel template synthesis. (a) In a polyacrylamide gel, the polymer chains form covalent crosslinks through N,N methylene-bis-acrylamide (circles). (b) Reaction of gelatin and methacrylic anhydride for grafting of methacryloyl substitution groups. The modification occurs at primary amine and hydroxyl groups. The RGD domains are presented as segments along the GelMA polymer chains, and their chemical structure is depicted within the inset. To create a hydrogel network, the methacrylated gelatin was crosslinked using ultraviolet light in the presence of a photoinitiator. (c) In a PA/GelMA hybrid gel, the two types of polymer network are intertwined and formed double networks. (d) Schematics of cell spheroid fabrication on PA/GelMA hydrogel template.

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Fig. 2

Characterization of PA/GelMA hydrogel composite. (a) SEM micrographs of hydrogels with varying compositions. (b) Quantification of pore size of different kinds of hydrogels. (c) Stress/strain curves of three kinds of hydrogels (PA, PA/5% w/v GelMA, and PA/10% w/v GelMA). Scale bar: 100 μm (up) and 50 μm (bottom).

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Fig. 3

Characterization of cell spheroids generated on PA/GelMA hydrogel templates. (a) Live/dead staining of MCF-7 breast cancer cells on PA, 5% w/v GelMA, PA/5% w/v GelMA, and PA/10% w/v GelMA hydrogel samples, respectively. Cell spheroids were well formed on both PA/5% w/v GelMA and PA/10% w/v GelMA hydrogel groups. (b) SEM micrographs of single cell spheroid on PA/10% w/v GelMA hydrogel templates at 1, 3, 7 days of culturing, respectively. (c) Diameter distribution of cell spheroids on PA/5% w/v GelMA and PA/10% w/v GelMA hydrogel templates. (d) Immunofluorescent staining of cell spheroids formed on PA/10% w/v GelMA hydrogel templates after 10 days of culturing. (F-actin (phalloidin); nuclei (DAPI)). (e,f) The influence of cell seeding density (e) and culturing time (f) on the diameter of cell spheroid formed on PA/10% w/v GelMA hydrogel templates. Scale bar: (a) 50 μm (b,d) 20 μm.

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