4.10. Differentiation Potential of Nano-Engineered MSCs

Nano-engineered MSCs were seeded at a density of ~4 × 10⁴ cells/well in a 24-well plate, and then incubated with adipogenic or osteogenic differentiation media (StemPro Osteogenesis or Adipogenesis Differentiation Kits, Life Technologies, Carlsbad, CA, USA) for 3 weeks. The medium was replaced every 3–4 days. The untreated MSCs incubated with differentiation media and those incubated with regular culture media were used as positive and negative controls, respectively. After a 3-week incubation period, cells were fixed with 4% formalin and stained with alizarin red or oil red O to visualize osteogenic and adipogenic differentiation, respectively. Images were captured using a light microscope at 20× magnification.