Supplementary Material for: Conditioned medium from human uterine cervical stem cells regulates oxidative stress and angiogenesis of retinal pigment epithelial cells.

Introduction: Retinal homeostasis is essential to avoid retinal pigment epithelium (RPE) damage resulting in photoreceptor death and blindness. Mesenchymal stem cells-based cell therapy could contribute to the maintenance of the retinal homeostasis. We have explored the effect of human uterine cervical stem cells (hUCESCs) conditioned medium (hUCESC-CM) on RPE cells under oxidative stress condition.
Methods: ARPE-19 cells were treated with hydrogen peroxide (H2O2) in presence or absence of hUCESC-CM. qRT-PCR and Western blot were used to evaluate the expression of oxidative stress (HO-1, GCLC and HSPB1) and vasculogenesis (VEGFA, PDGFA and PDGFB) related factors. Also, we assessed in vitro effects of hUCESC-CM on endothelial cells (HUVEC) tube formation.
Results: mRNA expression of HO-1, GCLC, HSPB1, VEGFA, PDGFA and PDGFB were significantly increased in ARPE-19 cells treated with H2O2 + hUCESC-CM compared to cells treated with H2O2 only. Regarding the tube formation assay, HUVEC treated with supernatant from ARPE-19 cells treated with H2O2 + hUCESC-CM showed a significant increase in average vessel length, number of capillary-like junctions and average of vessels area compared with HUVEC treated with supernatant from ARPE-19 cells treated with H2O2 only.
Conclusion: Our results show potential therapeutic effects of hUCESC-CM on RPE, such as protection from damage by oxidative stress, stimulation of detoxifying genes and a better vascularization.