SARS-CoV-2 Virus Detection Via a Polymeric Nanochannel-Based Electrochemical Biosensor.

The development of simple, cost-effective, rapid, and quantitative diagnostic tools remains critical to monitor infectious COVID-19 disease. Although numerous diagnostic platforms, including rapid antigen tests, are developed and used, they suffer from limited accuracy, especially when tested with asymptomatic patients. Here, a unique approach to fabricate a nanochannel-based electrochemical biosensor that can detect the entire virion instead of virus fragments, is demonstrated. The sensing platform has uniform nanoscale channels created by the convective assembly of polystyrene (PS) beads on gold electrodes. The PS beads are then functionalized with bioreceptors while the gold surface is endowed with anti-fouling properties. When added to the biosensor, SARS-CoV-2 virus particles block the nanochannels by specific binding to the bioreceptors. The nanochannel blockage hinders the diffusion of a redox probe; and thus, allows quantification of the viral load by measuring the changes in the oxidation current before and after virus incubation. The biosensor shows a low limit of detection of ≈1.0 viral particle mL-1 with a wide detection range up to 108 particles mL-1 in cell culture media. Moreover, the biosensor is able to differentiate saliva samples with SARS-CoV-2 from those without, demonstrating the potential of this technology for translation into a point-of-care biosensor product.

The Requirement of Genetic Diagnostic Technologies for Environmental Surveillance of Antimicrobial Resistance.

Antimicrobial resistance (AMR) is threatening modern medicine. While the primary cost of AMR is paid in the healthcare domain, the agricultural and environmental domains are also reservoirs of resistant microorganisms and hence perpetual sources of AMR infections in humans. Consequently, the World Health Organisation and other international agencies are calling for surveillance of AMR in all three domains to guide intervention and risk reduction strategies. Technologies for detecting AMR that have been developed for healthcare settings are not immediately transferable to environmental and agricultural settings, and limited dialogue between the domains has hampered opportunities for cross-fertilisation to develop modified or new technologies. In this feature, we discuss the limitations of currently available AMR sensing technologies used in the clinic for sensing in other environments, and what is required to overcome these limitations.

Deposition of Aminomalononitrile-Based Films: Kinetics, Chemistry, and Morphology.

In the last few years, the development of versatile coating chemistries has become a hot topic in surface science after the discovery that catecholamines can lead to conformal coatings upon oxidation from aqueous solutions. Recently, it was found that aminomalononitrile (AMN), a molecule implicated in the appearance of life on earth, is an excellent prototype of novel material-independent surface functionalizing agents leading to conformal and biocompatible coatings in a simple and direct chemical process from aqueous solutions. So far, very little insight has been gained regarding the mechanisms underlying coating deposition. In this paper, we show that the chemical evolution of AMN film deposition under slightly basic conditions is different in solution and on silica. Thereon, the coating proceeds via a nucleation process followed by further deposition of islands which evolve to produce nitrogen-rich superhydrophilic fibrillar structures. Additionally, we show that AMN-based material can form films at the air-solution interface from unshaken solutions. These results open new vistas into the chemistry of HCN-derived species of potential relevance in materials science.

Astrochemistry and Astrobiology: Materials Sciencein Wonderland?

Astrochemistry and astrobiology, the fascinating disciplines that strive to unravel the origin of life, have opened unprecedented and unpredicted vistas into exotic compounds as well as extreme or complex reaction conditions of potential relevance for a broad variety of applications. Representative, and so far little explored sources of inspiration include complex organic systems, such as polycyclic aromatic hydrocarbons (PAHs) and their derivatives; hydrogen cyanide (HCN) and formamide (HCONH2) oligomers and polymers, like aminomalononitrile (AMN)-derived species; and exotic processes, such as solid-state photoreactions on mineral surfaces, phosphorylation by minerals, cold ice irradiation and proton bombardment, and thermal transformations in fumaroles. In addition, meteorites and minerals like forsterite, which dominate dust chemistry in the interstellar medium, may open new avenues for the discovery of innovative catalytic processes and unconventional methodologies. The aim of this review was to offer concise and inspiring, rather than comprehensive, examples of astrochemistry-related materials and systems that may be of relevance in areas such as surface functionalization, nanostructures, and hybrid material design, and for innovative technological solutions. The potential of computational methods to predict new properties from spectroscopic data and to assess plausible reaction pathways on both kinetic and thermodynamic grounds has also been highlighted.

Controlled Attachment of Pseudomonas aeruginosa with Binary Colloidal Crystal-Based Topographies.

Micro- and nanotopographies can interfere with bacteria attachment, however, the interplay existing between surface chemistry and topography remains unclear. Here, self-assembled spherical micrometer- silica and nanometer poly(methyl methacrylate) (PMMA)-sized particles are used to make binary colloidal crystal (BCC) topographical patterns to study bacterial attachment. A uniform surface chemistry of allylamine plasma polymer (AAMpp) is coated on the top of the BCCs to study only the topography effects. The uncoated and coated BCCs are exposed to Pseudomonas aeruginosa, and the surfaces and bacteria are characterized using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and fluorescence microscopy. It is found that bacteria attachment to the uncoated BCCs is delayed and individual cells are attracted to the small particle regions of the patterns. Surprisingly, this phenomenon is also observed for the AAMpp-coated BCCs, with bacteria attaching to the small particle regions of the pattern, in stark contrast to uniform flat films of AAMpp that are highly adhesive toward P. aeruginosa. Also, the overall levels of bacterial attachment are significantly reduced by the BCC patterns compared to controls. Thus, there is a trade-off that exists between chemistry and topography that can be exploited to delay the onset of P. aeruginosa biofilm formation on surfaces.

Development of Silicone Hydrogel Antimicrobial Contact Lenses with Mel4 Peptide Coating.

SIGNIFICANCE: This study investigated the development of an antimicrobial coating on silicone hydrogel contact lenses that may have the capacity to reduce contact lens-related infection and inflammatory events. PURPOSE: The purpose of this study was to develop an effective antimicrobial coating for silicone hydrogel contact lenses by attachment of Mel4 peptide. METHODS: Lotrafilcon A, comfilcon A, somofilcon A, senofilcon A, and lotrafilcon B silicone hydrogel contact lenses were plasma coated with acrylic acid followed by Mel4 antimicrobial peptide immobilization by covalent coupling. Peptide immobilization was quantified by x-ray electron spectroscopy. Contact lens diameter, base curve, center thickness, and lens surface wettability were measured by captive-bubble contact-angle technique. Antimicrobial activity of the lenses was determined against Pseudomonas aeruginosa and Staphylococcus aureus by viable plate count and also after soaking with artificial tears solution for 1 day. In vivo safety and biocompatibility were determined in an animal model for 1 week. RESULTS: Mel4 peptide-coated silicone hydrogel contact lenses were associated with high antimicrobial inhibition (>2 log), except for lotrafilcon B and senofilcon A. Lotrafilcon B did not exhibit any activity, whereas senofilcon A showed 1.4- and 0.7-log inhibition against P. aeruginosa and S. aureus, respectively. X-ray electron spectroscopy revealed significant increases in the lens surface-bound amide nitrogen in all contact lenses except for lotrafilcon B. All contact lens parameters remained unchanged except for the base curve and center thickness for senofilcon A. Mel4 immobilization was associated with a decrease in contact angle. Mel4-coated contact lens wear was not associated with any signs or symptoms of ocular irritation in a rabbit model study. Reduced antimicrobial activity was observed with all the lenses after soaking with artificial tears solution or rabbit wear. CONCLUSIONS: Mel4 antimicrobial coating may be an effective option for development of antimicrobial silicone hydrogel contact lenses.

Modulation of human mesenchymal and pluripotent stem cell behavior using biophysical and biochemical cues: A review.

In vitro manipulation of human stem cells is a critical process in regenerative medicine and cellular therapies. Strategies and methods to maintain stem cells and direct them into specific lineages are ongoing challenges in these fields. To date, a number of studies have reported that besides biochemical stimulation, biophysical cues in the form of surface patterning and external stimulation also influence stem cell attachment, proliferation, and differentiation, and can be used in cell reprogramming and the maintenance of pluripotency. While biochemical cues are generally effective and easy to deliver, biophysical cues have many other advantages for scalability as they are cost efficient, have a longer lifetime, and can be easily defined. However, different protocols and cell sources utilized in a variety of studies have led to difficulties in obtaining clear conclusions about the effects of the biophysical environment on stem cells. In addition, the examination of different types of external stimulation is time consuming and limited by available fabrication techniques, resulting in a delay in commercialization and clinical applications. In this review, we aim to summarize the most important biophysical cues and methods for the culture of human stem cells, including mesenchymal and pluripotent stem cells, to facilitate their adoption in stem cell biology. The standard classical protocols of using biochemical cues will also be discussed for comparison. We believe that combining biochemical and biophysical stimulation has the greatest potential to generate functionally mature cells at a scalable and inexpensive rate for diverse applications in regenerative medicine and cell therapy. Biotechnol. Bioeng. 2017;114: 260-280. © 2016 Wiley Periodicals, Inc.

Facile One-step Micropatterning Using Photodegradable Methacrylated Gelatin Hydrogels for Improved Cardiomyocyte Organization and Alignment.

Hydrogels are often employed as temporary platforms for cell proliferation and tissue organization in vitro. Researchers have incorporated photodegradable moieties into synthetic polymeric hydrogels as a means of achieving spatiotemporal control over material properties. In this study protein-based photodegradable hydrogels composed of methacrylated gelatin (GelMA) and a crosslinker containing o-nitrobenzyl ester groups have been developed. The hydrogels are able to degrade rapidly and specifically in response to UV light and can be photopatterned to a variety of shapes and dimensions in a one-step process. Micropatterned photodegradable hydrogels are shown to improve cell distribution, alignment and beating regularity of cultured neonatal rat cardiomyocytes. Overall this work introduces a new class of photodegradable hydrogel based on natural and biofunctional polymers as cell culture substrates for improving cellular organization and function.

Enhanced tumour cell nuclear targeting in a tumour progression model.

BACKGROUND: There is an urgent need for new approaches to deliver bioactive molecules to cancer cells efficiently and specifically. METHODS: Here we fuse the cancer cell nuclear targeting module of the Chicken Anaemia Virus Apoptin protein to the core histones H2B and H3 and utilise them in transfection, protein transduction and DNA binding assays. RESULTS: We found subsequent nuclear accumulation of these proteins to be 2-3 fold higher in tumour compared to normal cells in transfected isogenic human osteosarcoma and breast tumour progression models. This represents the first demonstration of enhanced nuclear targeting by Apoptin in a tumour progression model, and its functionality in a heterologous protein context. Excitingly, we found that the innate transduction ability of histones could be exploited in combination with the Apoptin nuclear targeting module to effect an overall 13-fold higher delivery of protein to osteosarcoma cancer cell nuclei compared to their isogenic normal counterparts. CONCLUSIONS: This is the first report of cancer-cell specificity by a cell penetrating protein, with important implications for the use of protein transduction as a vehicle for gene/drug delivery in the future, and in particular in the development of highly specific and effective anti-cancer agents.

Photodegradable Gelatin-Based Hydrogels Prepared by Bioorthogonal Click Chemistry for Cell Encapsulation and Release.

In this study, we present a method for the fabrication of in situ forming gelatin and poly(ethylene glycol)-based hydrogels utilizing bioorthogonal, strain-promoted alkyne-azide cycloaddition as the cross-linking reaction. By incorporating nitrobenzyl moieties within the network structure, these hydrogels can be designed to be degradable upon irradiation with low intensity UV light, allowing precise photopatterning. Fibroblast cells encapsulated within these hydrogels were viable at 14 days and could be readily harvested using a light trigger. Potential applications of this new class of injectable hydrogel include its use as a 3D culturing platform that allows the capture and release of cells, as well as light-triggered cell delivery in regenerative medicine.

Patterning and biofunctionalization of antifouling hyperbranched polyglycerol coatings.

We demonstrate the patterned biofunctionalization of antifouling hyperbranched polyglycerol (HPG) coatings on silicon and glass substrates. The ultralow fouling HPG coatings afforded straightforward chemical handles for rapid bioconjugation of amine containing biomolecular species. This was achieved by sodium periodate oxidation of terminal HPG diols to yield reactive aldehyde groups. Patterned microprinting of sodium periodate and cell adhesion mediating cyclic peptides containing the RGD sequence resulted in an array of covalently immobilized bioactive signals. When incubated with mouse fibroblasts, the HPG background resisted cell attachment whereas high density cell attachment was observed on the peptide spots, resulting in high-contrast cell microarrays. We also demonstrated single-step, in situ functionalization of the HPG coatings by printing periodate and peptide concurrently. Our results demonstrate the effectiveness of antifouling and functionalized HPG graft polymer coatings and establish their use in microarray applications for the first time.

Polymerizable peptide copolymer coatings for the control of biointerfacial interactions.

The effective control over biointerfacial interactions is essential for a broad range of biomedical applications in vitro and in vivo such as biosensors, cell culture tools and implantable devices. Here, our aim was to develop a coating strategy that is transferable between different substrate materials and can effectively suppress nonspecific protein adsorption and hence reduce cell attachment while also presenting bioactive signals to enable specific cell-material interactions. In a first step an allylamine plasma polymer coating was applied, followed by the covalent immobilization of a macroinitiator carrying iniferter functionalities in the side chains. Subsequently, copolymers with different molar ratios of acrylamide and a polymerizable peptide containing the sequence Arg-Gly-Asp (RGD) were grafted via surface initiated free radical polymerization. X-ray photoelectron spectroscopy (XPS) was used to confirm the success of each coating step. The cellular response to these coatings was evaluated using L929 mouse fibroblast cell culture assays for up to 24 h. Cell attachment was significantly reduced on acrylamide homopolymer coatings and negative control surfaces representing a polymerizable peptide containing the nonbioactive Arg-Ala-Asp (RAD) sequence. In contrast, cell attachment was increased with increasing polymerizable RGD peptide ratios in the copolymer. The combination of acrylamide-terminated peptide sequences in combination with acrylamide provides a simple and versatile route to surfaces that combine low nonspecific protein adsorption and the display of controlled densities of bioactive signals and is expected to be translated into a number of biomedical applications in vitro and in vivo.

Inhibition of protein and cell attachment on materials generated from N-(2-hydroxypropyl) acrylamide.

Effective control over biointerfacial interactions is essential for a broad range of biomedical applications. At this point in time, only a relatively small range of radically polymerizable monomers have been described that are able to generate low fouling polymer materials and surfaces. The most important examples that have been successfully used in the context of the reduction of nonspecific protein adsorption and subsequent cell attachment include PEG-based monomers such as poly(ethylene glycol) methacrylate (PEGMA), zwitterionic monomers such as 2-methacryloyloxyethyl phosphorylcholine and noncharged monomers such as acrylamide and N-(2-hydroxypropyl) methacrylamide (HPMAm). However, issues such as oxidative degradation and poor polymerization characteristics limit the applicability of most of these candidates. Here we have synthesized the monomer N-(2-hydroxypropyl) acrylamide (HPAm), examined its polymerization kinetics and evaluated its suitability for RAFT mediated polymerization in comparison to HPMAm. We also synthesized hydrogels using HPMAm and HPAm and evaluated the ability of HPAm polymers to occlude protein adsorption and cell attachment. In RAFT-controlled polymerization, much faster (8×) polymerization was observed for HPAm relative to HPMAm and better control was achieved over the molecular weight distribution. The performance of hydrogels prepared from HPAm in the prevention of protein adsorption and cellular attachment was equivalent to or better than that observed for materials made from HPMAm and PEG. These results open the door for HPAm based polymers in applications where effective control over biointerfacial interactions is required.

Tackling catheter-associated urinary tract infections with next-generation antimicrobial technologies.

Urinary catheters and other medical devices associated with the urinary tract such as stents are major contributors to nosocomial urinary tract infections (UTIs) as they provide an access path for pathogens to enter the bladder. Considering that catheter-associated urinary tract infections (CAUTIs) account for approximately 75% of UTIs and that UTIs represent the most common type of healthcare-associated infections, novel anti-infective device technologies are urgently required. The rapid rise of antimicrobial resistance in the context of CAUTIs further highlights the importance of such preventative strategies. In this review, the risk factors for pathogen colonization in the urinary tract are dissected, taking into account the nature and mechanistics of this unique environment. Moreover, the most promising next-generation preventative strategies are critically assessed, focusing in particular on anti-infective surface coatings. Finally, emerging approaches in this field and their likely clinical impact are examined.

A surface-independent bioglue using photo-crosslinkable benzophenone moiety.

Surface coating technology is broadly demanded across various fields, including marine and biomedical materials; therefore, a facile and versatile approach is desired. This study proposed an attractive surface coating strategy using photo-crosslinkable benzophenone (BP) moiety for biomaterials application. BP-containing "bioglue" polymer can effectively crosslink with all kinds of surfaces and biomolecules. Upon exposure to ultraviolet (UV) light, free radical reaction from the BP glue facilitates the immobilization of diverse molecules onto different substrates in a straightforward and user-friendly manner. Through either one-step, mixing the bioglue with targeted biomolecules, or two-step methods, pre-coating the bioglue and then adding targeted biomolecules, polyacrylic acid (PAA), cyclic RGD-containing peptides, and proteins (gelatin, collagen, and fibronectin) were successfully immobilized on substrates. After drying the bioglue, targeted biomolecules can still be immobilized on the surfaces preserving their bioactivity. Cell culture on biomolecule-immobilized surfaces using NIH 3T3 fibroblasts and human bone marrow stem cells (hBMSCs) showed significant improvement of cell adhesion and activity compared to the unmodified control in serum-free media after 24 hours. Furthermore, hBMSCs on the fibronectin-immobilized surface showed an increased calcium deposition after 21 days of osteogenic differentiation, suggesting that the immobilized fibronectin is highly bioactive. Given the straightforward protocol and substrate-independent bioglue, the proposed coating strategy is promising in broad-range fields.

Layer-by-layer polymer coating on discrete particles of cubic lyotropic liquid crystalline dispersions (cubosomes).

Cubic phase lyotropic liquid crystalline colloidal dispersions (cubosomes) were surface-modified with seven polyelectrolyte layers using a layer-by-layer (LbL) approach. The first layer consisted of a copolymer synthesized from methacrylic acid and oleoyl methacrylate for enhanced incorporation within the bilayer of the cubic nanostructure. Six additional layers of poly(L-lysine) and poly(methacrylic acid) were then sequentially added, followed by a washing procedure to remove polymer aggregates from the soft matter particles. Polymer buildup was monitored via microelectrophoresis, dynamic light scattering, and small-angle X-ray scattering. Polymer-coated cubosomes were observed with cryo-transmission electron microscopy. A potential application of the modified nanostructured particles presented in this study is to reduce the burst-release effect associated with drug-loaded cubosomes. The effectiveness of this approach was demonstrated through loading and release results from a model hydrophilic small molecule (fluorescein).

Anti-infective characteristics of a new Carbothane ventricular assist device driveline.

Objectives: Driveline infections are a major complication of ventricular assist device (VAD) therapy. A newly introduced Carbothane driveline has preliminarily demonstrated anti-infective potential against driveline infections. This study aimed to comprehensively assess the anti-biofilm capability of the Carbothane driveline and explore its physicochemical characteristics. Methods: We assessed the Carbothane driveline against biofilm formation of leading microorganisms causing VAD driveline infections, including Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Candida albicans, using novel in vitro biofilm assays mimicking different infection micro-environments. The importance of physicochemical properties of the Carbothane driveline in microorganism-device interactions were analyzed, particularly focusing on the surface chemistry. The role of micro-gaps in driveline tunnels on biofilm migration was also examined. Results: All organisms were able to attach to the smooth and velour sections of the Carbothane driveline. Early microbial adherence, at least for S. aureus and S. epidermidis, did not proceed to the formation of mature biofilms in a drip-flow biofilm reactor mimicking the driveline exit site environment. The presence of a driveline tunnel however, promoted staphylococcal biofilm formation on the Carbothane driveline. Physicochemical analysis of the Carbothane driveline revealed surface characteristics that may have contributed to its anti-biofilm activity, such as the aliphatic nature of its surface. The presence of micro-gaps in the tunnel facilitated biofilm migration of the studied bacterial species. Conclusion: This study provides experimental evidence to support the anti-biofilm activity of the Carbothane driveline and uncovered specific physicochemical features that may explain its ability to inhibit biofilm formation.

The influence of dysfunctional actin on polystyrene-nanotube-mediated mRNA nanoinjection into mammalian cells.

The advancement of nanofabrication technologies has transformed the landscape of engineered nano-bio interfaces, especially with vertically aligned nanoneedles (NNs). This enables scientists to venture into new territories, widening NN applications into increasingly more complex cellular manipulation and interrogation. Specifically, for intracellular delivery application, NNs have been shown to mediate the delivery of various bioactive cargos into a wide range of cells-a physical method termed "nanoinjection". Silicon (Si) nanostructures demonstrated great potential in nanoinjection, whereas the use of polymeric NNs for nanoinjection has rarely been explored. Furthermore, the underlying mechanism of interaction at the cell-NN interface is subtle and multifaceted, and not fully understood-underpinned by the design versatility of the NN biointerface. Recent studies have suggested that actin dynamic plays a pivotal role influencing the delivery efficacy. In this study, we fabricated a new class of NNs-a programmable polymeric nanotubes (NTs)-from polystyrene (PS) cell cultureware, designed to facilitate mRNA delivery into mouse embryonic fibroblast GPE86 cells. The PSNT delivery platform was able to mediate mRNA delivery with high delivery efficiency (∼83%). We also investigated the role of actin cytoskeleton in PSNTs mediated intracellular delivery by introducing two actin inhibitors-cytochalasin D (Cyto D) and jasplakinolide (Jas)-to cause dysfunctional cytoskeleton, via inhibiting actin polymerization and depolymerization, respectively (before and after the establishment of cell-PSNT interface). By inhibiting actin dynamics 12 h before cell-PSNT interfacing (pre-interface treatment), the mRNA delivery efficiencies were significantly reduced to ∼3% for Cyto D-treated samples and ∼1% for Jas-treated sample, as compared to their post-interface (2 h after cell-PSNT interfacing) counterpart (∼46% and ∼68%, respectively). The added flexibility of PSNTs have shown to help withstand mechanical breakage stemming from cytoskeletal forces in contrast to the SiNTs. Such findings will step-change our capacity to use programmable polymeric NTs in fundamental cellular processes related to intracellular delivery.

Surface Characteristics and Bone Biocompatibility of Cold-Sprayed Porous Titanium on Polydimethylsiloxane Substrates.

A variant of the cold spray (CS) technique was applied for the functionalization of polymer-based materials such as polydimethylsiloxane (PDMS) to improve the extent of mammalian cell interactions with these substrates. This was demonstrated by the embedment of porous titanium (pTi) into PDMS substrates using a single-step CS technique. CS processing parameters such as gas pressure and temperature were optimized to achieve the mechanical interlocking of pTi in the compressed PDMS to fabricate a unique hierarchical morphology possessing micro-roughness. As evidenced by the preserved porous structure, the pTi particles did not undergo any significant plastic deformation upon impact with the polymer substrate. The thickness of the particle embedment layer was determined, by cross-sectional analysis, ranging from 120 µm to over 200 µm. The behavior of osteoblast-like cells MG63 coming into contact with the pTi-embedded PDMS was examined. The results showed that the pTi-embedded PDMS samples promoted 80-96% of cell adhesion and proliferation during the early stages of incubation. The low cytotoxicity of the pTi-embedded PDMS was confirmed, with cell viability of the MG63 cells being above 90%. Furthermore, the pTi-embedded PDMS facilitated the production of alkaline phosphatase and calcium deposition in the MG63 cells, as demonstrated by the higher amount of alkaline phosphatase (2.6 times) and calcium (10.6 times) on the pTi-embedded PDMS sample fabricated at 250 °C, 3 MPa. Overall, the work demonstrated that the CS process provided flexibility in the parameters used for the production of the modified PDMS substrates and is highly efficient for the fabrication of coated polymer products. The results obtained in this study suggest that a tailorable porous and rough architecture could be achieved that promoted osteoblast function, indicating that the method has promise in the design of titanium-polymer composite materials applied to biomaterials used in musculoskeletal applications.

Cell Microencapsulation within Gelatin-PEG Microgels Using a Simple Pipet Tip-Based Device.

Microgels are microscale particles of hydrogel that can be laden with cells and used to create macroporous tissue constructs. Their ability to support cell-ECM and cell-cell interactions, along with the high levels of nutrient and metabolite exchange facilitated by their high surface area-to-volume ratio, means that they are attracting increasing attention for a variety of tissue regeneration applications. Here, we present methods for fabricating and modifying the structure of microfluidic devices using commonly available laboratory consumables including pipet tips and PTFE and silicon tubing to produce microgels. Different microfluidic devices realized the controlled generation of a wide size range (130-800 µm) of microgels for cell encapsulation. Subsequently, we describe the process of encapsulating mesenchymal stromal cells in microgels formed by photo-cross-linking of gelatin-norbornene and PEG dithiol. The introduced pipet-based chip offers simplicity, tunability, and versatility, making it easily assembled in most laboratories to effectively produce cell-laden microgels for various applications in tissue engineering.