Effect of adipose-derived stem cells-conditioned medium extracellular vesicles on senescent fibroblast and E2F1 expression

Authors

  • Vidya Hana Dwi Ayuningtyas Departement of Dermatology and Venereology, Faculty of Medicine, Universitas Brawijaya, dr. Saiful Anwar Hospital, Malang, Indonesia
  • Suci Prawitasari Departement of Dermatology and Venereology, Faculty of Medicine, Universitas Brawijaya, dr. Saiful Anwar Hospital, Malang, Indonesia
  • Aunur Rofiq Departement of Dermatology and Venereology, Faculty of Medicine, Universitas Brawijaya, dr. Saiful Anwar Hospital, Malang, Indonesia
  • Sinta Murlistyarini Departement of Dermatology and Venereology, Faculty of Medicine, Universitas Brawijaya, dr. Saiful Anwar Hospital, Malang, Indonesia

DOI:

https://doi.org/10.13181/mji.oa.247217

Keywords:

adipose-derived stem cells, extracellular vesicles, E2F1, senescence

Abstract

BACKGROUND Adipose-derived stem cells (ADSCs) are well-known for their regenerative properties, especially towards senescent cells. Extracellular vesicles derived from ADSCs are believed to influence the expression level of the E2 promoter binding factor (E2F1) protein, one of the key proteins regulating the cell cycle. This study aimed to investigate the impact of extracellular vesicles from ADSC-conditioned medium (ADSC-CM) on E2F1 levels and their potential to improve aging cells.

METHODS Extracellular vesicles from ADSC-CM were introduced into senescent fibroblasts through transfection. Then, the E2F1 protein levels were measured and compared between transfected and untransfected cells. A total of 18 samples were calculated based on Federer’s formulas. E2F1 protein levels were counted using a cell-based enzyme-linked immunosorbent assay. Senescence-associated beta-galactosidase staining was used to quantify the number of senescent cells in each group, and the microculture tetrazolium technique assay was used to assess cellular metabolic activity.

RESULTS The number of senescent cells was lower in the transfected group compared to the untransfected group. ADSC-CM extracellular vesicles-transfected fibroblasts exhibited higher levels of E2F1 protein (0.19 [0.17] ng/ml) compared to untransfected fibroblast (0.06 [0.049] ng/ml; p = 0.048). Higher E2F1 protein levels were associated with reduced senescent fibroblasts and increased metabolic viable fibroblasts in the transfected group.

CONCLUSIONS ADSC-CM extracellular vesicles positively affected senescent cells by enhancing the level of E2F1.

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References

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Huang YC, Lai LC. The potential roles of stem cell-derived extracellular vesicles as a therapeutic tool. Ann Transl Med. 2019;7(22):693. https://doi.org/10.21037/atm.2019.11.66

Mens MM, Ghanbari M. Cell cycle regulation of stem cells by microRNAs. Stem Cell Rev Rep. 2018;14(3):309−22. https://doi.org/10.1007/s12015-018-9808-y

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Murlistyarini S, Aninda LP, Widyarti S, Endharti AT, Sardjono TW. Exosomes of adipose-derived stem cells promotes retinoblastoma and forkhead-box M1 protein expression. Maced J Med Sci. 2021;9(A):422-7. https://doi.org/10.3889/oamjms.2021.6195

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Kishore J, Schumacher SM, Srikanth GV, Koch WJ. Exosomes from G coupled receptor kinase (GRK2) knock-down cells suppress fibroblast apoptosis potential mechanism by which GRK2 inhibition provides protection after myocardial infarction. Int J Sci Eng Res. 2016;7(7): 229-36. https://doi.org/10.1126/scitranslmed.aaa0154

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Noren Hooten N, Evans MK. Techniques to induce and quantify cellular senescence. J Vis Exp. 2017;(123):55533. https://doi.org/10.3791/55533

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Giglio S, Cirombella R, Amodeo R, Portaro L, Lavra L, Vecchione A. Micro RNA miR-24 promotes cell proliferation by targeting the CDKs inhibitors p27Kip1 and p16INK4a. J Cell Physiol. 2013;228(10):2015-23. https://doi.org/10.1002/jcp.24368

Oh M, Lee J, Kim YJ, Rhee WJ, Park JH. Exosomes derived from human induced pluripotent stem cells ameliorate the aging of skin fibroblasts. Int J Mol Sci. 2018;19(6):1715. https://doi.org/10.3390/ijms19061715

Tominaga T, Shimada R, Okada Y, Kawamata T, Kibayashi K. Senescence-associated-β-galactosidase staining following traumatic brain injury in the mouse cerebrum. PLoS One. 2019;14(3):e0213673. https://doi.org/10.1371/journal.pone.0213673

Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, et al. Cell viability assays. 2013 [updated 2016 Jul 1]. In: Markossian S, Grossman A, Arkin M, Auld D, Austin C, Baell J, et al. Assay guidance manual [Internet]. Bethesda: Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-.

Published

2024-12-03

How to Cite

1.
Ayuningtyas VHD, Prawitasari S, Rofiq A, Murlistyarini S. Effect of adipose-derived stem cells-conditioned medium extracellular vesicles on senescent fibroblast and E2F1 expression. Med J Indones [Internet]. 2024Dec.3 [cited 2024Dec.5];33(3):141-7. Available from: https://mji.ui.ac.id/journal/index.php/mji/article/view/7217

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Section

Basic Medical Research
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