| 研究実績の概要 |
At present, the cellular senescence of the mesenchymal stem cells (MSCs) on various stiffness substrates was investigated by in vitro expansion of the cells on hydrogels in comparison with tissue culture dish. The senescence state was elucidated by quantification of cell, nuclear size and immunofluorescence staining of CSK related proteins. The senescence cells were identified using senescence associated beta-galactosidase (SA-beta-GAL) staining. The cell motility was analyzed by time-lapse observation. The expression of senescence markers such as galactosidase beta-1 (GLB1), and cell cycle inhibitors, p16, p21 and p53 were monitored using real-time PCR.
Serially passaged the MSCs on both soft (3 kPa) and stiff (20 kPa) hydrogels decreased the expression of GLB1, p16, p21 p53 and decreased the numbers of SA-beta-GAL positive staining in comparison to the tissue culture dish. On the hydrogels, size of the cells and nucleus were decreased, and their migration properties could also be rescued.
These results suggested that the expansion of the MSCs on hydrogel substrates could reverse the aging phenotypes and delayed senescence.
|
