Fluorescence Imaging of Actin Turnover Parses Early Stem Cell Lineage Divergence and Senescence.

ABSTRACT: This study describes a new approach to discern early divergence in stem cell lineage progression via temporal dynamics of the cytoskeletal protein, F-actin. The approach involves real-time labeling of human mesenchymal stem cells (MSCs) and longitudinal tracking of the turnover dynamics of a fluorogenic F-actin specific probe, SiR-actin (SA). Cells cultured in media with distinct lineage factors and labeled with SA showed lineage specific reduction in the actin turnover shortly after adipogenic (few minutes) and chondrogenic (3-4 hours) commitment in contrast to osteogenic and basal cultured conditions. Next, composite staining of SA along with the competing F-actin specific fluorescent conjugate, phalloidin, and high-content image analysis of the complementary labels showed clear phenotyp

SUBMITTER: Mishra P 

PROVIDER: S-EPMC6637207 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature