Removal from adherent culture contributes to apoptosis in human bone marrow mesenchymal stem cells

Mesenchymal stem cells (MSCs) are routinely isolated due to their adherence to tissue culture plates and their in vitro growth characteristics. Expansion of MSCs in adherent cultures is the only way to obtain sufficient cells for use in either clinical or research settings. MSCs have tremendous potential in myocardial repair treatment by cell therapy techniques, however, a large number of MSCs die from apoptosis following transplantation. Previous studies have examined the factors contributing to the survival of transplanted cells, but little is known about the effect of removal from adherent culture conditions on apoptosis of the MSCs. In the present study, human bone marrow MSCs were expanded in adherent cultures. Then apoptosis rates were examined at different time points in MSCs cultured in nonadherent conditions (ultra-low-adherence plates) compared with MSCs cultured in adherent conditions (standard tissue culture plates). Flow cytometry analysis suggested that cell apoptosis increased when MSCs were cultured in nonadherent culture conditions. In addition, western blot and reverse transcription-quantitative polymerase chain reaction analyses demonstrated that caspase-3, −7 and −9 were involved in this process. The present study demonstrated that loss of culture adherence increases apoptosis of human MSCs. The present findings may provide new insight into the factors affecting MSC survival after transplantation.