Gi Seok Jeong

Digitally tunable physicochemical coding of material composition and topography in continuous microfibres

Heterotypic functional materials with compositional and topographical properties that vary spatiotemporally on the micro- or nanoscale are common in nature. However, fabricating such complex materials in the laboratory remains challenging. Here we describe a method to continuously create microfibres with tunable morphological, structural and chemical features using a microfluidic system consisting of a digital, programmable flow control that mimics the silk-spinning process of spiders. With this method we fabricated hydrogel microfibres coded with varying chemical composition and topography along the fibre, including gas micro-bubbles as well as nanoporous spindle-knots and joints that enabled directional water collection. We also explored the potential use of the coded microfibres for tissue engineering applications by creating multifunctional microfibres with a spatially controlled co-culture of encapsulated cells.

solderable and electroplatable flexible electronic circuit on a porous stretchable elastomer

A variety of flexible and stretchable electronics have been reported for use in flexible electronic devices or biomedical applications. The practical and wider application of such flexible electronics has been limited because commercial electronic components are difficult to be directly integrated into flexible stretchable electronics and electroplating is still challenging. Here, we propose a novel method for fabricating flexible and stretchable electronic devices using a porous elastomeric substrate. Pressurized steam was applied to an uncured polydimethylsiloxane layer for the simple and cost-effective production of porous structure. An electroplated nickel anchor had a key role in bonding commercial electronic components on elastomers by soldering techniques, and metals could be stably patterned and electroplated for practical uses. The proposed technology was applied to develop a plaster electrocardiogram dry electrode and multi-channel microelectrodes that could be used as a long-term wearable biosignal monitor and for brain signal monitoring, respectively.

Applications of micromixing technology

This review presents an application of micromixer technologies, which have driven a number of critical research trends over the past few decades, particularly for chemical and biological fields. Micromixer technologies in this review are categorized according to their applications: (1) chemical applications, including chemical synthesis, polymerization, and extraction; (2) biological applications, including DNA analysis, biological screening enzyme assays, protein folding; and (3) detection/analysis of chemical or biochemical content combined with NMR, FTIR, or Raman spectroscopies. In the chemical application, crystallization, extraction, polymerization, and organic synthesis have been reported, not only for laboratory studies, but also for industrial applications. Microscale techniques are used in chemical synthesis to develop microreactors. In clinical medicine and biological studies, microfluidic systems have been widely applied to the identification of biochemical products, diagnosis, drug discovery, and investigation of disease symptoms. The biological and biochemical applications also include enzyme assays, biological screening assays, cell lysis, protein folding, and biological analytical assays. Nondestructive analytical/detection methods have yielded a number of benefits to chemical and biochemical processes. In this chapter, we introduce analytical methods those are frequently integrated into micromixing technologies, such as NMR, FT-IR, and Raman spectroscopies. From the study of micromixers, we discovered that the Re number and mixing time depends on the specific application, and we clustered micromixers in various applications according to the Re number and mixing performance (mixing time). We expect that this clustering will be helpful in designing of micromixers for specific applications.

Cell encapsulation via microtechnologies

The encapsulation of living cells in a variety of soft polymers or hydrogels is important, particularly, for the rehabilitation of functional tissues capable of repairing or replacing damaged organs. Cellular encapsulation segregates cells from the surrounding tissue to protect the implanted cell from the recipients immune system after transplantation. Diverse hydrogel membranes have been popularly used as encapsulating materials and permit the diffusion of gas, nutrients, wastes and therapeutic products smoothly. This review describes a variety of methods that have been developed to achieve cellular encapsulation using microscale platform. Microtechnologies have been adopted to precisely control the encapsulated cell number, size and shape of a cell-laden polymer structure. We provide a brief overview of recent microtechnology-based cell encapsulation methods, with a detailed description of the relevant processes. Finally, we discuss the current challenges and future directions likely to be taken by cell microencapsulation approaches toward tissue engineering and cell therapy applications.

Sprouting angiogenesis under a chemical gradient regulated by interactions with an endothelial monolayer in a microfluidic platform.

Microfluidic cell culture assays are versatile tools for studying cell migration, particularly angiogenesis. Such assays can deliver precisely controlled linear gradients of chemical stimuli to cultured cells in a microfluidic channel, offering excellent optical resolution and in situ monitoring of cellular morphogenesis in response to a gradient. Microfluidic cell culture assays provide a chemical gradient subject to molecular diffusion, although cellular metabolism can perturb it. The actual gradient perturbed by cells has not been precisely described in the context of regulated cellular morphogenesis. We modeled the chemical gradient in a microfluidic channel by simulating the analyte(VEGF) distribution during cellular interactions. The results were experimentally verified by monitoring sprouting angiogenic response from a monolayer of human umbilical vein endothelial cells (hUVECs) into a type 1 collagen scaffold. The simulation provided a basis for understanding a real distribution of the analyte interrupted by cells in microfluidic device. The new protocol enables one to quantify the morphogenesis of hUVECs under a flat, less-steep, or steep gradient.

Surface Tension-Mediated, Concave-Microwell Arrays for Large-Scale, Simultaneous Production of Homogeneously Sized Embryoid Bodies

Embryonic stem cells (ESCs) are pluripotent and capable of self-renewal. ESC aggregates, termed embryoid bodies (EBs), have been widely adopted as an in vitro differentiation model. However, the mass production of uniform size and shaped EBs has been challenging. Herein is described the development of a culture plate containing a large number of concave microwells with minimal use of tools, labor, skill, and cost, enabling the production of a large number of homogeneous EBs simultaneously using the culture plate. The large number of concave well structures is self-constructed through the surface tension of the viscoelastic PDMS prepolymer. Murine ESCs (mESCs) are then seeded onto the concave wells for mass production of monodisperse EBs. It is observed that the EBs produced over a large area are uniform in shape and size regardless of microwell position and differences in cell seeding densities, and whether their phenotype is maintained. The capability to differentiate into adult cells (neuron and endothelial cells) from EBs is also evaluated and the neural spikes from differentiated neuron cells are measured to observe their function. Uniform size and shape EBs are successfully generated in large scale and their pluripotency is maintained similar to other methods.

Microfluidic assay of endothelial cell migration in 3D interpenetrating polymer semi-network HA-Collagen hydrogel

Cell migration through the extracellular matrix (ECM) is one of the key features for physiological and pathological processes such as angiogenesis, cancer metastasis, and wound healing. In particular, the quantitative assay of endothelial cell migration under the well-defined three dimensional (3D) microenvironment is important to analyze the angiogenesis mechanism. In this study, we report a microfluidic assay of endothelial cell sprouting and migration into an interpenetrating polymer semi-network HA-Collagen (SIPNs CH) hydrogel as ECM providing an enhanced in vivo mimicking 3D microenvironment to cells. The microfluidic chip could provide a well-controlled gradient of growth factor to cells, whereas the hydrogel could mimic a well-defined 3D microenvironment in vivo. (In addition/Furthermore, the microfluidic chip gives a well-controlled gradient of growth factor to cells) For this reason, three types of hydrogel, composed of semi-interpenetrating networks of collagen and hyaluronic acid were prepared, and firstly we proved the role of the hydrogel in endothelial cell migration. The diffusion property and swelling ratio of the hydrogel were characterized. It modulated the migration of endothelial cells in quantified manner, also being influenced by additional synthesis of Matrix metalloproteinase(MMP)-sensitive remodeling peptides and Arginine–glycine–lycinee (RGD) cell adhesion peptides. We successfully established a novel cell migration platform by changing major determinants such as ECM material under biochemical synthesis and under growth factor gradients in a microfluidic manner.

Large-Scale, Ultrapliable, and Free-Standing Nanomembranes

The creation and characterization of large-area ultrathin highly pliable free-standing PDMS membranes and their application to the study of cellular epithelia is described. The ultra-thin membranes permitted the straight forward calculation of cell monolayer moduli, derived from measured stress-strain curves. These measurements allowed the unprecedented detection of cellular-level injury in the epithelia caused by the rupture of cell-cell tight junctions in response to stretching.

Networked neural spheroid by neuro-bundle mimicking nervous system created by topology effect

In most animals, the nervous system consists of the central nervous system (CNS) and the peripheral nervous system (PNS), the latter of which connects the CNS to all parts of the body. Damage and/or malfunction of the nervous system causes serious pathologies, including neurodegenerative disorders, spinal cord injury, and Alzheimer’s disease. Thus, not surprising, considerable research effort, both in vivo and in vitro, has been devoted to studying the nervous system and signal transmission through it. However, conventional in vitro cell culture systems do not enable control over diverse aspects of the neural microenvironment. Moreover, formation of certain nervous system growth patterns in vitro remains a challenge. In this study, we developed a deep hemispherical, microchannel-networked, concave array system and applied it to generate three-dimensional nerve-like neural bundles. The deep hemicylindrical channel network was easily fabricated by exploiting the meniscus induced by the surface tension of a liquid poly(dimethylsiloxane) (PDMS) prepolymer. Neurospheroids spontaneously aggregated in each deep concave microwell and were networked to neighboring spheroids through the deep hemicylindrical channel. Notably, two types of satellite spheroids also formed in deep hemispherical microchannels through self-aggregation and acted as an anchoring point to enhance formation of nerve-like networks with neighboring spheroids. During neural-network formation, neural progenitor cells successfully differentiated into glial and neuronal cells. These cells secreted laminin, forming an extracellular matrix around the host and satellite spheroids. Electrical stimuli were transmitted between networked neurospheroids in the resulting nerve-like neural bundle, as detected by imaging Ca2+ signals in responding cells.

Networked neuro-spheres formed by topological attractants for engineering of 3-dimensional nervous system

The nervous systems including central and peripheral nervous system have an important role in transmitting signals from brain to organs and tissues. Due to such critical function of nervous system, considerable effort has been tried to establish in vitro nervous model. In this paper, the neuro-spheres networked by the nerve-like structure were created using concave well arrays connected by the hemicylindrical channels. The concave microwells and the hemicylindrical channels were fabricated using the surface tension of viscose liquid PDMS prepolymer for the creation of neuro-spheres-networking (NSN). To investigate the topological effect of the concave-well-hemicylindrical-channel-networking (CWHCN), comparative experiments were conducted on a conventional cylindrical-wells-rectangular-channel-networking (CWRCN). Neuro-progenitor cells from the rat were seeded on the concave well arrays connected by the hemicylindrical channels and cultured for 10 days. Small neuro-pehroids consisting of neurons and glia cells were autonomously formed in the concave microwell arrays. These neuro-spheres were networked by the nerve-like structures formed along the CWHCN. In order to confirm the interconnection of the neurites in the NSN, a calcium imaging experiment was performed to measure the calcium flux along the nerve-like networking. To demonstrate further use of the networked neuro-networking for brain regeneration, we transferred the NSN onto hydrogel maintaining approximately 90% viability, and this model is expected to be used for the regeneration of a damaged brain.