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Myosin-X (MYO10) is an actin-based motor protein involved in cytoskeletal dynamics, membrane interactions, and integrin-mediated adhesion. To investigate MYO10's cellular roles, we generated MYO10 knockdown (MYO10 ) HeLa and COS7 cell lines using lentiviral shRNA. Compared to wild-type cells, both MYO10 lines showed reduced proliferation and impaired cell migration in wound assays. Additionally, there were fewer edge filopodia in HeLa cells. Furthermore, MYO10 cells demonstrated increased spreading on laminin-coated substrates, suggesting altered integrin activation and cytoskeletal linkage. Our results reinforce MYO10's importance in cell proliferation, adhesion, and migration. These MYO10 lines provide an accessible cell culture model for future study of MYO10.