The iMSCs utilized in this study were derived from induced pluripotent stem cells (iPSCs) (Nuwacell, Cat #RC1001, China), following the previously reported methodology [53 (link)]. To characterize the iMSCs, flow cytometry was employed to analyze the presence of typical MSC markers. The iMSCs were cultured in an incubator at 37 °C and 5% CO2 using MSC basal medium (Dakewe, China) supplemented with EliteGro™-Advanced serum-free supplement (Dakewe, China).
Doxorubicin (APExBIO, USA) was utilized to induce cell senescence in iMSCs. Specifically, iMSCs were cultured in MSC basal medium supplemented with serum-free supplement and 100 nM Doxorubicin for 2 days to obtain senescent iMSCs exhibiting altered morphology. Control samples consisted of iMSCs cultured in MSC basal medium supplemented with a serum-free supplement and PBS for 2 days.
Senolytics (ABT-263, NMN, and metformin; Absin, China) were employed to modulate the senescent phenotype of iMSC. Specifically, iMSCs (passages 12–14) were treated with these three drugs for 2 days each.
Doxorubicin (APExBIO, USA) was utilized to induce cell senescence in iMSCs. Specifically, iMSCs were cultured in MSC basal medium supplemented with serum-free supplement and 100 nM Doxorubicin for 2 days to obtain senescent iMSCs exhibiting altered morphology. Control samples consisted of iMSCs cultured in MSC basal medium supplemented with a serum-free supplement and PBS for 2 days.
Senolytics (ABT-263, NMN, and metformin; Absin, China) were employed to modulate the senescent phenotype of iMSC. Specifically, iMSCs (passages 12–14) were treated with these three drugs for 2 days each.