High-throughput physical phenotyping of cell differentiation

Abstract

In this report, we present multiparameter deformability cytometry (m-DC), in which we explore a large set of parameters describing the physical phenotypes of pluripotent cells and their derivatives. m-DC utilizes microfluidic inertial focusing and hydrodynamic stretching of single cells in conjunction with high-speed video recording to realize high-throughput characterization of over 20 different cell motion and morphology-derived parameters. Parameters extracted from videos include size, deformability, deformation kinetics, and morphology. We train support vector machines that provide evidence that these additional physical measurements improve classification of induced pluripotent stem cells, mesenchymal stem cells, neural stem cells, and their derivatives compared to size and deformability alone. In addition, we utilize visual interactive stochastic neighbor embedding to visually map the high-dimensional physical phenotypic spaces occupied by these stem cells and their progeny and the pathways traversed during differentiation. This report demonstrates the potential of m-DC for improving understanding of physical differences that arise as cells differentiate and identifying cell subpopulations in a label-free manner. Ultimately, such approaches could broaden our understanding of subtle changes in cell phenotypes and their roles in human biology.

Cells Under Stress: An Inertial-Shear Microfluidic Determination of Cell Behavior

“…Previous studies have shown that different microfluidic flow regimes can alter the mechanical response of cells. Gossett et al (14,17,18) developed the technique deformability cytometry (DC) (19,20) in which cells are hydrodynamically stretched at the stagnation point (SP) of an extensional flow device at rates up to 2000 cells/s. A compressional force (F C ) due to fluid inertia and a shear force (F S ) due to fluid viscosity act on the cells.…”

The mechanics of myeloid cells

“…The differentiation of progenitor cells is coupled to major changes in their mechanical properties. This is partially due to differences in intracellular organisation and composition [Yourek et al., ; Darling et al., ; Lin et al., ]. In some instances, progenitor mechanical properties can serve as predictive biomarkers for differentiation outcomes [González‐Cruz et al., ].…”

On-chip cell manipulation and applications to deformability measurements

“…In addition, more improvements in the user interface of microfluidic devices and software are also important requirements for the usage in the clinical situations by clinical doctors, not only by engineers in laboratories. In this paradigm, automation of both the hardware and software is another important topic to be addressed, and has been intensively investigated [91][92][93][94][95]. For the on-chip cell manipulation, we are also developing the automation of the gain-tuning toward clinical usages [96].…”