Wettability‐Based Cell Sorting: Exploring Label‐Free Isolation Strategy for Mixed Primary Glial Cell Population

Abstract

with antibodies against cell-type-specific membrane proteins that are conjugated to either magnet or fluorophores. Cell sorting instruments are then used to separate these labeled cells. [3] The labeling-based separation has an inherently high specificity by definition, ensuring good purification quality. However, the labeling-based methods have the disadvantages of high cost, technical complexity, and low yield. In addition, the labeling step during the isolation process may cause undesired changes in cellular properties. [2] Labelfree approaches, on the other hand, allow isolation of the target cells based on their physical properties such as size, density, deformability, and adhesiveness. [4,5] Since physical approaches to cell isolation provide greater accessibility and lower costs than biological approaches, cell separation methods based on the physical properties of cells are currently being widely used. Despite the better accessibility, the physical approach may be considered more delicate because its accuracy is critically dependent on the skill of individual researchers. Among different physical traits of cells, cellular adhesion properties, including cell-matrix adhesion and cell-cell adhesion, are considered the marked traits for identifying distinct cell types or physiological states. For example, in cell state classification, cells of mesenchymal phenotype mainly exhibit robust cell-matrix adhesion, whereas epithelial cells are characterized by prominent cell-cell adhesion. [6] In cancer cells, metastatic cells typically display relatively weaker cell-matrix adhesion than their non-metastatic counterparts. [7] Cell-matrix adhesion strength is also known to contribute to lineage determination during the differentiation of mesenchymal stem cells. [8] Accordingly, the adhesion strength has been utilized as a sorting strategy for cells of different origins and pathophysiological states.Glial cells, which are non-neuronal cells in the brain, have gained considerable attention as key players in neurological diseases. [9] Glial cell populations consist of three types of cells, namely astrocytes, oligodendrocytes, and microglia, each with distinct functions in the brain: astrocytes interact with neurons and blood vessels; [10] oligodendrocytes serve as the primary electrical insulator for neurons; [11] microglia are the critical immune regulators. [12] These three types of glial cells are derived from different origins, thereby exhibiting distinct Glial brain cells, including astrocytes, oligodendrocytes, and microglia, have received much attention as crucial players in neurological diseases. As a result of their critical roles, numerous in vitro studies are being conducted, necessitating the use of appropriate isolation methods for different glial cell types. The most effective glial isolation at the moment is labeling-based protocols that require expensive antibodies and equipment. More commonly used label-free methods are better known for higher accessibility but with compromised accuracy and longer isol...

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