Microfluidic Generation of Amino-Functionalized Hydrogel Microbeads Capable of On-Bead Bioassay

Kim, Seong-Soo; Lee, Sang-Myung; Lee, Sung-Sik; Shin, Dong‐Sik

doi:10.3390/mi10080527

Cited by 4 publications

(3 citation statements)

References 34 publications

(37 reference statements)

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“…They are inherently non-adhesive to cells and unless modified they are non-biodegradable. Functionalization with molecules such as Arg-Gly-Asp (RGD) motifs (Sionkowska, 2011;Kim et al, 2019), or mixing of natural and synthetic polymers (Krutkramelis et al, 2016), is therefore required for many cellular applications. Alterations of these polymers can be used to provide useful cross-linking approaches such as photopolymerization as demonstrated in PEG (Young et al, 2013;Xia et al, 2017) and PVA (Zhao et al, 2010).…”

Section: Synthetic Hydrogelsmentioning

confidence: 99%

Hydrogels for Single-Cell Microgel Production: Recent Advances and Applications

Tiemeijer

Tel

2022

Front. Bioeng. Biotechnol.

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Single-cell techniques have become more and more incorporated in cell biological research over the past decades. Various approaches have been proposed to isolate, culture, sort, and analyze individual cells to understand cellular heterogeneity, which is at the foundation of every systematic cellular response in the human body. Microfluidics is undoubtedly the most suitable method of manipulating cells, due to its small scale, high degree of control, and gentle nature toward vulnerable cells. More specifically, the technique of microfluidic droplet production has proven to provide reproducible single-cell encapsulation with high throughput. Various in-droplet applications have been explored, ranging from immunoassays, cytotoxicity assays, and single-cell sequencing. All rely on the theoretically unlimited throughput that can be achieved and the monodispersity of each individual droplet. To make these platforms more suitable for adherent cells or to maintain spatial control after de-emulsification, hydrogels can be included during droplet production to obtain “microgels.” Over the past years, a multitude of research has focused on the possibilities these can provide. Also, as the technique matures, it is becoming clear that it will result in advantages over conventional droplet approaches. In this review, we provide a comprehensive overview on how various types of hydrogels can be incorporated into different droplet-based approaches and provide novel and more robust analytic and screening applications. We will further focus on a wide range of recently published applications for microgels and how these can be applied in cell biological research at the single- to multicell scale.

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Section: Synthetic Hydrogelsmentioning

confidence: 99%

Hydrogels for Single-Cell Microgel Production: Recent Advances and Applications

Tiemeijer

Tel

2022

Front. Bioeng. Biotechnol.

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“…Two major advantages of hydrogels are that they provide relevant mechanical cues and allow for cell adherence ( Huang et al, 2017 ; Mohamed et al, 2019 ). Commonly used in droplet microfluidics are the natural occurring alginate-based ( Leong et al, 2016 ; Huang et al, 2017 ; Mao et al, 2019 ) or synthetic polyethylene glycol-based (PEG) ( Krutkramelis et al, 2016 ; Kim et al, 2019 ) hydrogels which are primarily gelated using chemical or physical crosslinking. Although these techniques are well developed and understood addition of gelation agents has the potential to negatively affect cell phenotype and viability.…”

Section: Introductionmentioning

confidence: 99%

Probing Single-Cell Macrophage Polarization and Heterogeneity Using Thermo-Reversible Hydrogels in Droplet-Based Microfluidics

Tiemeijer

Sweep

Sleeboom

et al. 2021

Front. Bioeng. Biotechnol.

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Human immune cells intrinsically exist as heterogenous populations. To understand cellular heterogeneity, both cell culture and analysis should be executed with single-cell resolution to eliminate juxtacrine and paracrine interactions, as these can lead to a homogenized cell response, obscuring unique cellular behavior. Droplet microfluidics has emerged as a potent tool to culture and stimulate single cells at high throughput. However, when studying adherent cells at single-cell level, it is imperative to provide a substrate for the cells to adhere to, as suspension culture conditions can negatively affect biological function and behavior. Therefore, we combined a droplet-based microfluidic platform with a thermo-reversible polyisocyanide (PIC) hydrogel, which allowed for robust droplet formation at low temperatures, whilst ensuring catalyzer-free droplet gelation and easy cell recovery after culture for downstream analysis. With this approach, we probed the heterogeneity of highly adherent human macrophages under both pro-inflammatory M1 and anti-inflammatory M2 polarization conditions. We showed that co-encapsulation of multiple cells enhanced cell polarization compared to single cells, indicating that cellular communication is a potent driver of macrophage polarization. Additionally, we highlight that culturing single macrophages in PIC hydrogel droplets displayed higher cell viability and enhanced M2 polarization compared to single macrophages cultured in suspension. Remarkably, combining phenotypical and functional analysis on single cultured macrophages revealed a subset of cells in a persistent M1 state, which were undetectable in conventional bulk cultures. Taken together, combining droplet-based microfluidics with hydrogels is a versatile and powerful tool to study the biological function of adherent cell types at single-cell resolution with high throughput.

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“…Nasser et al [4] demonstrated a PMMA-based microfluidics device for cost-effective PCR applications. Kim et al [5] made homogenous amino-functionalized hydrogel microbeads for on-bead bioassay.…”

mentioning

confidence: 99%