Anoikis-resistant OS are viable and exhibit lower growth rates. A-C: Representative AI-grown OS cells generated from mHOS (A), 143B (B), and a patient derived OS cell line, MS206-2 (C). D: Growth rates measured as cell doublings per day. Results shown are average ± standard error. Asterisks indicate statistical significance (**p < 0.01, ***p < 0.001). E: Western blot analysis of 143B and MG-63 OS cells shows decreased Ki67 protein levels in AI grown cells (P2 spheres) compared to adherent cells.