2023 Volume 37 Issue 2 Pages 88-95
Mesenchymal stem cells (MSCs) have the potential to differentiate into endothelial cells (ECs) and become an attractive cell source for the application of therapeutic regenerative medicine. For developing highly efficient differentiation induction, we investigated the effect of a combination of biomechanical cyclic stretching and spheroid culture on MSC differentiation into endothelial cells. Spheroids were fabricated with adipose-derived stem cells (ADSCs), which were precultured with EC conditioned media and exposed to fluid shear stress for 3 days. After being cultured statically for 4 days in polydimethylsiloxane stretching chambers, the spheroids were exposed to 10% cyclic stretching at 1 Hz for 3 days. We confirmed that the spheroids maintained adhesion to the stretching chambers coated with 0.3 mg/ml type I collagen under the cyclic stretching condition. Meanwhile, the spheroids exhibited an elongated shape in response to the stretching. Real-time PCR analysis showed that the application of cyclic stretching to the ADSC spheroids induced EC-specific gene expression and hindered the other cell-type marks compared to the statically cultured spheroids. These results indicate that the combination of cyclic stretching and spheroid culture has the potential to efficiently induce EC differentiation of MSCs.
