Progestin Repairs the Mitochondria Membrane Potential by Preventing Membrane Hyperpolarization in Mitochondria Transferred Endometrial Stromal Cells

Background: The regenerative capacity of the endometrium is attributed to stem/progenitor cells. Despite their remarkable regenerative capacity, in some cases, impairments in regeneration can be observed. Endometrial mitochondria transplantation into the uterine cavity improves the uterine environment in Asherman’s syndrome. Mitochondria transfer shows therapeutic advantages by supporting tissue metabolism and viability. However, the disruptive environment of the endometrium could affect mitochondrial health adversely. Increasing mitochondrial activity with progestin protects against apoptosis. The aim was to investigate the effect of progestin supplementation during exogenous mitochondrial transplantation. The study was designed as an in vitro cell culture of endometrial stromal cells. Mitochondria were isolated from the same cell line, representing autologous mitochondria transplantation. Material and Method: The transplantation of mitochondria was detected by fluorescence labeling of mitochondria. Viability was assessed by CCK8, and apoptosis was detected by AnnexinV/PI staining. Gene expression analysis was performed for Ki67, p38, and Erk1/2. Results: Mitochondria were successfully transferred into endometrial stromal cells. The viability was not significantly altered due to the exogenous mitochondria, despite the increase in the reactive oxygen level. The addition of progestin is also well tolerated. The combined application of both mitochondria and progestin further supports the viability of cells without inducing the level of reactive oxygen species. Apoptotic levels were decreased in the presence of progestin even in the co-transfection group of mitochondria and progestin. The mitochondrial cell membrane was evaluated with JC-1 showing that the disrupted membrane potential was recovered by progestin, improving the damaged membrane potential of the mitochondria. Conclusion: The damaged membrane potential improved in the presence of progestin, helping to improve the overall output of the mitochondrial transplantation.