Single-Cell-Derived Tumor-Sphere Formation and Drug-Resistance Assay Using an Integrated Microfluidics

Abstract

Considerable evidence points to cancer stem-like cells (CSCs) as responsible for promoting progression, metastasis, and drug resistance. Without damage to the cell biological properties, single-cell-derived tumor-sphere is encouraging options for CSCs identification and studies. Although several single cell-based microfluidic methods have been developed for CSCs studies, clarifying liaison between the biomechanics of cells (such as size and deformability) and stem (such as tumor-sphere formation and drug resistance) remains challenging. Herein, we present a platform of integrated microfluidics for the analysis of single-cell-derived tumor-sphere formation and drug resistance. Tumor-spheres derived from different biomechanics (size and/or deformation) single-cells could be formed efficiently using this device. To demonstrate the microfluidic-platform capability, a proof-of-concept experiment was implemented by evaluating single-cell-derived sphere formation of single glioblastoma cells with different biomechanics. Additionally, a course of chemotherapy to study these single-cell-derived spheres was determined by coculture with vincristine. The results indicate that tumor cell biomechanics is associated with single-cell-derived spheres formation; that is, smaller and/or more deformable tumor cells are more stem-like defined by the formation of single-cell-derived spheres than more prominent and/or lesser deformable tumor cells. Also, tumor-spheres derived from single small and/or more deformable tumor cell have higher drug resistance than more prominent and/or less deformable tumor cells. Our device offers a new approach for single-cell-derived sphere formation according to tumor cell different biomechanical properties. Furthermore, it offers a new method for CSC identification and downstream analysis on a single-cell level.

Microfluidics-Based Single-Cell Research for Intercellular Interaction

“…The cellular heterogeneity are commonly identified at the phenotypic, transcriptomic or genomic levels (Treutlein et al, 2014;Pang et al, 2015Pang et al, , 2016Khoo et al, 2018;Lawson et al, 2018). Populations of multicellular constructs exhibit heterogeneities, total organizational and morphology changes have been demonstrated to simulate tumors or organs complexity (Lecault et al, 2011;Pang et al, 2019a). Microfluidics-based single-cell analysis methods have become a powerful tool to delve into the cell microenvironment, including immune cell studies.…”

PC3-Secreted Microprotein Is Expressed in Glioblastoma Stem-Like Cells and Human Glioma Tissues

“…11,12) Therefore, one promising approach to isolate CSCs is considered to form single-cell-derived tumorsphere because aggregation of anchorage-dependent cells must be prevented. 13,14) In fact, this approach succeeded in the establishment of CSC lines from human colon cancer cell line, 15) human primary colorectal cancer, 16) human liver cancer, 17) and human glioblastoma. 18) In this study, we established single cell-derived tumorspheres from human glioblastoma cell line U87MG, and one of the established tumorsphere, P4E8, showed CSC-like properties.…”

Global Trends of Organoid and Organ-On-a-Chip in the Past Decade: A Bibliometric and Comparative Study

“…And these 50 common documents contained 24 review articles and 26 research articles. Specifically, these research articles were related to the topics, including drug toxicology, 14 biomaterials, 15 organogenesis, 16 and 3D printing. 17,18 Besides, Lab on a Chip (n = 4) was ranked first in the source journals of these 50 documents.…”