Fabrication of Complex Hydrogel Materials by Utilizing Microfluidics and Micromolding
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Fabrication of Complex Hydrogel Materials by Utilizing Microfluidics and Micromolding Masumi Yamada, Sari Sugaya, Aoi Kobayashi, Ayaki Miyama, Yoji Naganuma, Emi Yamada, Shunta Kakegawa, and Minoru Seki Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, Chiba University, Chiba 263-8522, Japan.
ABSTRACT Here, we present microfluidic methods to fabricate complex hydrogel structures for 3D tissue or organ-like cell structures in vitro. First, a microfluidic system to continuously synthesize chemically and physically anisotropic Ca–alginate hydrogel microfibers is proposed to enable the guidance of cell proliferation and differentiation. Next, the microfluidic preparation methods for yarn-ball-shape hydrogel particles and extremely-small hydrogel microspheres. Finally, a newly developed micro-molding and bonding method for hydrogel micro-patterned plates is reported. INTRODUCTION A number of trials for in vitro construction of three-dimensional tissues or organ-like structures have been conducted, and various types of approaches have been developed by many researchers. A rapid in vitro assembly of cells to form complicated tissue-like structures composed of different cell patterns and embedded with vascular networks requires the precise, high-density, and three dimensional arrangements of cell-containing unit structures. In addition, the physicochemical properties of the microenvironment surrounding the cells should be controlled. However, development of a practical methodology for 3D cell assemblies has just been embarked upon. In recent years, several methods have been reported on producing micrometer-size cell-laden objects with highly-controlled morphologies and compositions by using the flow stream in microfabricated channels. Here we present microfluidic devices for preparing functional micrometer-size hydrogel materials having fibrous or particulate morphology. The physical/chemical heterogeneity of the prepared materials allows the incorporated cells to grow differently from the conventional plate cultivation, which is useful for preparing unit structures mimicking the in vivo tissues. In addition to these materials, here we introduce recently developed several microfluidic and microfabrication techniques, including the preparation of yarn-ball-shaped hydrogel structures [1], production of extremely-small hydrogel microspheres [2], preparation processes of microstructured and layered hydrogel plates [3], and micropatterning of ultra-thin hydrogels utilizing local surface modification. In addition, micronozzle structures for producing actuating lipid vesicles and continuous microfluidic cell sorting/treatment systems are presented. EXPERIMENTAL Preparation of anisotropic hydrogel fibers
We first prepared physically and chemically anisotropic Ca-alginate hydrogel microfibers by using axisymmetric microchannels. Sodium alginate (NaA), propylene glycol alginate (PGA), and calcium chloride were used to prepare alginate hydrogel fibers. Fluorescent microbeads with different diameters
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