A Novel Single-Step Fabrication Technique to Create Heterogeneous Poly(ethylene glycol) Hydrogel Microstructures Containing Multiple Phenotypes of Mammalian Cells (original) (raw)

Microfluidics-based fabrication of cell-laden microgels

Biomicrofluidics, 2020

Microfluidic principles have been extensively utilized as powerful tools to fabricate controlled monodisperse cell-laden hydrogel microdroplets for various biological applications, especially tissue engineering. In this review, we report recent advances in microfluidic-based droplet fabrication and provide our rationale to justify the superiority of microfluidics-based techniques over other microtechnology methods in achieving the encapsulation of cells within hydrogels. The three main components of such a system—hydrogels, cells, and device configurations—are examined thoroughly. First, the characteristics of various types of hydrogels including natural and synthetic types, especially concerning cell encapsulation, are examined. This is followed by the elucidation of the reasoning behind choosing specific cells for encapsulation. Next, in addition to a detailed discussion of their respective droplet formation mechanisms, various device configurations including T-junctions, flow-foc...

Responsive Micromolds for Sequential Patterning of Hydrogel Microstructures

Journal of the American Chemical Society, 2011

Microscale hydrogels have been shown to be beneficial for various applications such as tissue engineering and drug delivery. A key aspect in these applications is the spatial organization of biological entities or chemical compounds within hydrogel microstructures. For this purpose, sequentially patterned microgels can be used to spatially organize either living materials to mimic biological complexity or multiple chemicals to design functional microparticles for drug delivery. Photolithographic methods are the most common way to pattern microscale hydrogels but are limited to photocrosslinkable polymers. So far, conventional micromolding approaches use static molds to fabricate structures, limiting the resulting shapes that can be generated. Herein, we describe a dynamic micromolding technique to fabricate sequentially patterned hydrogel microstructures by exploiting the thermoresponsiveness of poly(N-isopropylacrylamide)based micromolds. These responsive micromolds exhibited shape changes under temperature variations, facilitating the sequential molding of microgels at two different temperatures. We fabricated multicompartmental striped, cylindrical, and cubic microgels that encapsulated fluorescent polymer microspheres or different cell types. These responsive micromolds can be used to immobilize living materials or chemicals into sequentially patterned hydrogel microstructures which may potentially be useful for a range of applications at the interface of chemistry, materials science and engineering, and biology.

Facile fabrication processes for hydrogel-based microfluidic devices made of natural biopolymers

Biomicrofluidics, 2014

We present facile strategies for the fabrication of two types of microfluidic devices made of hydrogels using the natural biopolymers, alginate, and gelatin as substrates. The processes presented include the molding-based preparation of hydrogel plates and their chemical bonding. To prepare calcium-alginate hydrogel microdevices, we suppressed the volume shrinkage of the alginate solution during gelation using propylene glycol alginate in the precursor solution along with sodium alginate. In addition, a chemical bonding method was developed using a polyelectrolyte membrane of poly-L-lysine as the electrostatic glue. To prepare gelatin-based microdevices, we used microbial transglutaminase to bond hydrogel plates chemically and to cross-link and stabilize the hydrogel matrix. As an application, mammalian cells (fibroblasts and vascular endothelial cells) were cultivated on the microchannel surface to form three-dimensional capillary-embedding tissue models for biological research and tissue engineering. V

Facile Microfluidic Fabrication of Biocompatible Hydrogel Microspheres in a Novel Microfluidic Device

Molecules

Poly(ethylene glycol) diacrylate (PEGDA) microgels with tuneable size and porosity find applications as extracellular matrix mimics for tissue-engineering scaffolds, biosensors, and drug carriers. Monodispersed PEGDA microgels were produced by modular droplet microfluidics using the dispersed phase with 49–99 wt% PEGDA, 1 wt% Darocur 2959, and 0–50 wt% water, while the continuous phase was 3.5 wt% silicone-based surfactant dissolved in silicone oil. Pure PEGDA droplets were fully cured within 60 s at the UV light intensity of 75 mW/cm2. The droplets with higher water content required more time for curing. Due to oxygen inhibition, the polymerisation started in the droplet centre and advanced towards the edge, leading to a temporary solid core/liquid shell morphology, confirmed by tracking the Brownian motion of fluorescent latex nanoparticles within a droplet. A volumetric shrinkage during polymerisation was 1–4% for pure PEGDA droplets and 20–32% for the droplets containing 10–40 w...

Biomimetic Dextran-Based Hydrogel Layers for Cell Micropatterning over Large Areas Using the FluidFM BOT Technology

Langmuir, 2019

Micropatterning of living single cells and cell clusters over millimeter-centimeter scale areas is of high demand in the development cell-based biosensors. Micropatterning methodologies require both a suitable biomimetic support and a printing technology. In this work we present the micropatterning of living mammalian cells on carboxymethyl dextran (CMD) hydrogel layers using the FluidFM BOT technology. In contrast to the ultrathin (few nanometers thick in dry state) CMD films generally used in label-free biosensor applications, we developed CMD layers with thicknesses of several tens of nanometers in order to provide support for the controlled adhesion of living cells. The fabrication method and detailed characterization of the CMD layers are also described. The antifouling ability of the CMD surfaces is demonstrated by in situ optical waveguide lightmode spectroscopy (OWLS) measurements using serum modelling proteins with different electrostatic properties and molecular weights. Cell micropatterning on the CMD surface was obtained by printing cell adhesion mediating cRGDfK peptide molecules (cyclo(Arg-Gly-Asp-D-Phe-Lys)) directly from aqueous solution using microchanneled cantilevers with subsequent incubation of the printed surfaces in living cell culture. Uniquely, we present cell patterns with different geometries (spot, line, grid arrays) covering both micrometer and millimeter-centimeter scale areas. The adhered patterns were analyzed by phase contrast microscopy and the adhesion process on the patterns was real-time monitored by digital holographic microscopy, enabling to quantify the survival and migration of cells on the printed cRGDfK arrays.

Control of Mammalian Cell and Bacteria Adhesion on Substrates Micropatterned with Poly(ethylene glycol) Hydrogels

Biomedical Microdevices, 2003

A simple method for controlling the spatial positioning of mammalian cells and bacteria on substrates using patterned poly(ethylene glycol) (PEG) hydrogel microstructures is described. These microstructures were fabricated using photolithography on silicon, glass or poly (dimethylsiloxane) (PDMS) surfaces modified with a 3-(trichlorosilyl) propyl methacrylate (TPM) monolayer. During the photogelation reaction, the resulting hydrogel microstructures were covalently bound to the substrate via the TPM monolayer and did not detached from the substrate upon hydration. For mammalian cell patterning, microwell arrays of different dimensions were fabricated. These microwells were composed of hydrophilic PEG hydrogel walls surrounding hydrophobic TPM floors inside the microwells. Murine 3T3 fibroblasts and transformed hepatocytes were shown to selectively adhere to the TPM monolayer inside the microwells, maintaining their viability, while adherent cells were not present on the hydrogel walls. The number of cells inside one microwell could be controled by changing the lateral dimension of the microwells, thus allowing only a single cell per microwell if desired. In the case of 30×30 μm microwells, as many as 400 microwells were fabricated in 1 mm2. In addition, PEG hydrogel microstructures were also shown to effectively resist the adhesion of bacteria such as Escherichia coli.

Hydrogels as artificial matrices for cell seeding in microfluidic devices

RSC Advances, 2020

Hydrogel-based artificial scaffolds play a vital role in shifting in vitro models from two-dimensional (2D) cell culture to three-dimensional (3D) cell culture. Microfluidic 3D cell culture systems with a hydrogel matrix encourage biomedical researchers to replace in vivo models with 3D in vitro models with a cellular microenvironment that resembles physiological conditions with greater fidelity. Hydrogels can be designed as an artificial extracellular matrix scaffold for providing spatial orientation and promoting cellular interactions with surroundings. Selecting the appropriate hydrogels and their fabrication techniques are the key to mimic the in vivo mechanical environment. Moreover, combining a microfluidic technique with a hydrogel-based 3D cell culture system can create a complex and controlled microenvironment for the cells by placing small biosamples inside the microchannel. This paper provides an overview of the structural similarities of the hydrogels as an extracellular matrix (ECM), their classification and fabrication techniques as an ECM, and their use in microfluidic 3D cell culture systems. Finally, the paper presents the current challenges and future perspectives of using hydrogel scaffolds in microfluidic 3D cell culture systems.

Generation of dynamic microenvironment in a hydrogel-based microfluidic device for cell culture study

2012

Many cellular characteristics such as cell attachment, migration, morphology and proliferation are affected by porosity and pore size of bioactive gradients. Therefore, engineering a gradient that mimics the porosity relevant features of different living tissues can provide a better understanding for some of these cellular characteristics. In this work, a simple method is proposed to generate an injectable hydrogel gradient that allows the pore structure to be altered post injection resulted in a continuous porosity scaffold gradient under. This gradient was employed to study the cell metastasis phenomenon through monitoring the chemotactic response of human fibrosarcoma (HT1080) cells, an invasive cancer cell model, in a cell culture housed in a polydimethylsiloxane (PDMS) microfluidic chip. It was found that the migration of cells through this gradient is sensitive to the mean pore size where cells migrated toward the largest pores. This gradient provides the advantage of giving u...

Hydrogels and microtechnologies for engineering the cellular microenvironment

Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology, 2011

Hydrogels represent a class of materials suitable for numerous biomedical applications such as tissue engineering and drug delivery. Hydrogels are by definition capable of absorbing large amount of fluid, making them adequate for cell seeding and encapsulation as well as for implantation because of their biocompatibility and excellent diffusion properties. They also possess other desirable properties for fundamental research as they have the ability to mimic the basic three-dimensional (3D) biological, chemical, and mechanical properties of native tissues. Furthermore, their biological interactions with cells can be modified through the numerous side groups of the polymeric chains. Thus, the biological, chemical, and mechanical properties, as well as the degradation kinetics of hydrogels can be tailored depending on the application. In addition, their fabrication process can be combined with microtechnologies to enable precise control of cell-scale features such as surface topography and the presence of adhesion motifs on the hydrogel material. This ability to control the microscale structure of hydrogels has been used to engineer tissue models and to study cell behavior mechanisms in vitro. New approaches such as bottom-up and directed assembly of microscale hydrogels (microgels) are currently emerging as powerful methods to enable the fabrication of 3D constructs replicating the microenvironment found in vivo. of cell-laden microgels for fabrication of 3D tissue constructs.