We detail the creation and function of a microfluidic device, which employs a passive, geometric method to effectively trap individual DNA molecules in chambers, enabling the detection of tumor-specific biomarkers.
Collecting circulating tumor cells (CTCs), target cells that are non-invasively obtained, is essential to biological and medical research. Complex procedures are frequently employed for conventional cell collection, entailing either size-differentiated sorting or invasive enzymatic reactions. This study showcases the development of a functional polymer film, comprising thermoresponsive poly(N-isopropylacrylamide) and conductive poly(34-ethylenedioxythiopene)/poly(styrene sulfonate), and its application for the capture and release of circulating tumor cells. The proposed polymer films, when coated onto microfabricated gold electrodes, possess the ability to capture and control the release of cells in a noninvasive manner, concurrently facilitating the monitoring of these processes through conventional electrical measurements.
Through the application of stereolithography based additive manufacturing (3D printing), novel in vitro microfluidic platforms are being created and developed. This manufacturing approach results in decreased production time, coupled with the ability to rapidly refine designs and create complex, solid structures. This chapter's platform is dedicated to capturing and evaluating cancer spheroids within a perfusion system. Spheroids, prepared in 3D Petri dishes, are stained and then carefully introduced into 3D printed imaging devices, where imaging is performed under continuous flow conditions. In contrast to traditional static monolayer cultures, this design supports active perfusion, leading to longer viability within complex 3D cellular constructs and improved in vivo condition mimicking results.
From inhibiting cancer growth by releasing pro-inflammatory compounds to aiding in its progression by secreting growth factors, immunomodulatory agents, and matrix-modifying enzymes, immune cells play a substantial role in the overall cancer process. Thus, the ex vivo analysis of immune cells' secretion processes can be utilized as a dependable prognostic biomarker for cancer cases. However, a noteworthy impediment in existing techniques for assessing the ex vivo secretion of cells is their low throughput and the requirement for a large volume of samples. The integration of cell culture and biosensors within a monolithic microdevice, a hallmark of microfluidics, grants a distinct benefit; it enhances analytical throughput while capitalizing on the inherent low-sample requirement. Additionally, the presence of fluid control elements promotes the automation of this analysis, leading to more reliable and consistent outcomes. We delineate a method for assessing the ex vivo secretory capacity of immune cells, utilizing a sophisticated, integrated microfluidic platform.
Circulating tumor cell (CTC) clusters, exceptionally rare and found in the bloodstream, can be isolated for minimally invasive diagnostic and prognostic purposes, revealing their contribution to metastasis. Innovations explicitly designed to enrich CTC clusters often fail to achieve the necessary processing throughput required in clinical settings, or they introduce detrimental high shear forces due to their design, putting large clusters at risk of damage. microfluidic biochips This methodology details a process for quickly and effectively isolating CTC clusters from cancer patients, irrespective of cluster size or cell surface markers. Cancer screening and personalized medicine will fundamentally incorporate the minimally invasive access to tumor cells found within the hematogenous circulation.
Small extracellular vesicles (sEVs), nanoscopic bioparticles, serve as a mode of intercellular transport for biomolecular cargoes. Several pathological conditions, including cancer, are linked to the use of electric vehicles, making them potentially valuable targets for therapeutic and diagnostic tools. Analyzing variations in the sEV biomolecular cargo's makeup may illuminate how these vesicles contribute to cancer. However, accomplishing this is made challenging by the analogous physical characteristics of sEVs and the crucial need for highly sensitive analytical processes. The sEV subpopulation characterization platform (ESCP), a platform using surface-enhanced Raman scattering (SERS) readouts for a microfluidic immunoassay, is detailed in our method of preparation and operation. To enhance the collisions of sEVs with the antibody-functionalized sensor surface, ESCP employs an electrohydrodynamic flow induced by an alternating current. in vivo immunogenicity For multiplexed and highly sensitive phenotypic characterization of captured sEVs, plasmonic nanoparticles are used for labeling, leveraging SERS. ESCP analysis reveals the expression levels of three tetraspanins (CD9, CD63, CD81) and four cancer-associated biomarkers (MCSP, MCAM, ErbB3, LNGFR) within sEVs isolated from cancer cell lines and plasma samples.
Blood and other bodily fluids are examined in liquid biopsies to determine the classification of malignant cells. Far less intrusive than tissue biopsies, liquid biopsies entail merely a small volume of blood or bodily fluids sampled from the patient. By utilizing microfluidics, researchers can isolate cancer cells from fluid biopsies, enabling early diagnosis of cancer. Microfluidic devices are finding an expanding application in the ever-evolving field of 3D printing. 3D printing surpasses conventional microfluidic device manufacturing in numerous aspects, including the seamless mass production of exact copies, the integration of diverse materials, and the accomplishment of complex or lengthy processes not easily achievable through microfluidic techniques. Enfortumab vedotin-ejfv compound library chemical Microfluidic chips augmented by 3D printing provide a relatively inexpensive platform for analyzing liquid biopsies, offering advantages over conventional microfluidic designs. In this chapter, we will dissect a 3D microfluidic chip-based method for separating cancer cells from liquid biopsies using affinity, as well as its underlying justifications.
Strategies for anticipating the efficacy of a given treatment for a particular patient are becoming a growing focus within the field of oncology. With its precision, personalized oncology holds the potential for a substantial lengthening of a patient's survival time. Therapy testing in personalized oncology relies predominantly on patient-derived organoids as a source of patient tumor tissue. Standard multi-well plates, coated with Matrigel, are the gold standard method for cancer organoid culture. While these standard organoid cultures are effective, they suffer from limitations: a large initial cell count is required, and the sizes of the resulting cancer organoids exhibit significant variation. The following deficiency hinders the monitoring and quantification of organoid size adjustments in relation to therapy. Microfluidic devices incorporating microwell arrays offer a means to decrease the initial cellular quantity required for organoid development, while simultaneously ensuring consistent organoid sizes, leading to streamlined therapy evaluations. We provide the methods for designing and developing microfluidic devices, and for introducing patient-derived cancer cells, cultivating organoids, and testing treatment strategies within these systems.
Rare circulating tumor cells (CTCs), present in the bloodstream in small numbers, serve as indicators of cancer progression. Although highly purified, intact CTCs with desired viability are crucial, their scarcity amidst blood cells presents a significant obstacle. The fabrication and application procedures for a novel self-amplified inertial-focused (SAIF) microfluidic chip, designed for size-based, high-throughput, label-free separation of circulating tumor cells (CTCs) from patient blood, are detailed in this chapter. The SAIF chip, introduced in this chapter, effectively demonstrates the viability of a highly narrow, zigzag channel (40 meters wide), connected to expansion regions, for efficiently separating cells of disparate sizes, maximizing their separation distance.
Pleural effusions containing malignant tumor cells (MTCs) signal the presence of malignancy. Despite this, the precision of MTC identification is considerably lowered by the overwhelming presence of background blood cells in large-scale specimens. An integrated system, combining an inertial microfluidic sorter and an inertial microfluidic concentrator, provides a method for the on-chip separation and enrichment of malignant pleural tumor cells (MTCs) from malignant pleural effusions (MPEs). By employing intrinsic hydrodynamic forces, the designed sorter and concentrator precisely guide cells to their designated equilibrium positions. This enables the sorting of cells based on size and the removal of cell-free fluids, resulting in enriched cell populations. This method leads to a 99.9% elimination of background cells and a tremendously high, nearly 1400-fold, enrichment of MTCs from significant amounts of MPEs. Immunofluorescence staining of the concentrated, high-purity MTC solution directly facilitates precise MPE identification, utilizing its high purity. Rare cell detection and quantification in various clinical samples can also be accomplished using the suggested approach.
Extracellular vesicles, exosomes, play a crucial role in intercellular communication between cells. Their accessibility across body fluids, including blood, semen, breast milk, saliva, and urine, alongside their bioavailability, has prompted their consideration as a non-invasive diagnostic, monitoring, and prognostic tool for various diseases, including cancer. Exosomes, when isolated and subsequently analyzed, are proving a valuable tool in diagnostic and personalized medicine. Despite its widespread adoption, the isolation procedure of differential ultracentrifugation is nonetheless arduous, time-consuming, expensive, and ultimately results in a restricted yield. Innovative microfluidic devices are emerging as platforms for exosome isolation, a low-cost technique that offers high purity and rapid treatment procedures.