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

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 isolation time. In this study, a cost-effective surface wettability-based isolation protocol for mixed glial cell population that does not require labeling is established. Specifically, when cultured on a hydrophobized substrate coated with a thin-film of poly (cyclohexyl methacrylate)(pCHMA), glial cells spontaneously segregate into substrate-adherent microglia and spheroids of astrocyte/oligodendrocyte progenitor cells in suspension. In comparison to existing label-free isolation methods, the strategic approach ensures rapid isolation and high purity. Furthermore, this is the first to demonstrate that the hydrophobic property of pCHMA-coated substrate can be used to sort multiple brain glial cell types based on their differential adhesion properties without the need for complex labeling procedure. The study emphasizes the wettability-controlled substrate's potential for segregating different cell types of various pathophysiological states.