Role of Antioxidant Components on Formation of 3D Human Embryonic Stem Cell-Derived Organoids

Background: According to Eiraku protocol, the human embry-onic stem cells (hESCs, RH6 cell line) colonies were dissociat-ed to single cells using accumax enzyme and plated in U-shape low adherent 98 well plate.Materials and Methods: In addition of Rock inhibitor, Y-27632 which was considered as control group, different an-tioxidant components including Tempol, Alpha lipoic Acid, Ascorbic Acid, Quercetin and N-acetylcysteine with different concentrations were used.. Two days later, the cells survival was assessed under an inverted microscope. Also, the aggregate size was analyzed one week later.Results: Our data expressed that high concentrations of Quercetin and tempol induced the cell death of hESCs, even in the presence of Y-27632. Whereas the lower concentrations of these components play a protective role in dissociated HESCs. The measurement of aggregate size confirmed the protective ef-fect of different antioxidant components to increase the quality of hESC derived organoids.Conclusion: Taking advantage of the original retina organoid protocol, we investigated the effects of antioxidant compo-nents after dissociation of human embryonic stem cells. Our results showed that although some high concentrations of antioxidant revealed some deleterious effects on cell viability, in almost all optimal concentration of antioxidants, the survival was improved comparing control group. Therefore, one of the future efforts will be to apply and optimize the antioxidant component(s) to the human retinal organoid system.

کلیدواژه ها:

Antioxidant ، Organoid ، Embryonic Stem Cell