Adipose-derived stem cells (ASCs) therapies are emerging as a promising approach to therapeutic regeneration. Therapeutic persistence and reduced functional stem cell following cell delivery remain a critical hurdle for the clinical investigation due to the senescence of freshly isolated ASCs, extensive in-vitro passage. Cultured ASCs were derived from subcutaneous white adipose tissue isolated from mice fed a normal diet (NFD) or high-fat fed (HFD). ASCs were passaged in vitro and prepared for subculture up to passage 9, after light and non-light-treated ASCs. We performed senescence-associated-β-galactosidase (SA-β-gal) staining and real-time PCR to evaluate the time-dependent changes in expression levels of mRNA related to cell cycle, cellular senescence markers, and senescence-associated secretory phenotype (SASP). The mRNA expression levels of senescence and SASP markers were significantly increased at passages 7 and 9 of ASCs-NFD. At the same passage number, ASCs-HFD showed a markedly higher mRNA level of senescence and SASP than that in ASCs-NFD. At different passages (Passages 3, 5, 7, and 9), these cells were performed by light treatment. Photomodulation reduced the expression of senescence, SASP, and SA-β-Gal in all cells at passage. Moreover, the light-treated ASCs-derived exosomes decrease the expression of senescence, SASP, and SA-β-Gal in the later passage cells. Furthermore, Light treatment significantly increased the Ca 2 + influx compared to the non-light-treated ASCs. Collectively, ASCs can undergo cellular senescence in-vitro passage. Photomodulation may be better preserved over senescence in aged ASCs. Light-treated ASCs-derived exosomes could be served as e a new protective paradigm for the cellular senescence in-vitro passage.