Student The Catholic University of Korea, Republic of Korea
Abstract: Mitochondria play an important role in cell metabolism and survival, producing chemical energy through oxidative phosphorylation. When mitochondria are damaged, normal cells transfer their mitochondria to dysfunctional cells, promoting their revitalization. For this reason, its potential application in the treatment of damaged tissues has been widely investigated. Mitochondria derived from induced pluripotent stem cells (iPSCs) are expected to exhibit superior therapeutic effects due to their pluripotent nature. The mitochondria of iPSCs are distinguished by their low metabolic rate and reduced reactive oxygen species (ROS) production while generating sufficient ATP. Therefore, we aim to develop iPSC-derived mitochondria therapy for osteoporosis (OP), where ROS play a critical role in pathogenesis, and first confirmed its effect on in vitro osteogenesis. This study focuses on isolating mitochondria from iPSCs and delivering them directly to cells. We first isolated mitochondria from iPSCs using a commercial mitochondria isolation kit and validated their presence and purity using Western blot and TEM imaging. The quality of isolated donor mitochondria is important for mitochondria transfer therapy efficiency. Therefore, we checked the mitochondrial membrane potential and ATP levels of the isolated donor mitochondria to assess the quality of them. Next, we observed internalized donor mitochondria in the recipient cell after transfer by confocal microscopy, and we also determined senescence of the recipient iPSC after mitochondria transfer. After checking mitochondrial quality and mitochondria transfer validity, we transferred the isolated mitochondria to iPSCs prior to osteogenic differentiation and observed changes in their differentiation capacity. This proof-of-concept study demonstrates the potential of iPSC-derived mitochondria transfer to be applied to in vitro bone formation, providing a basis for the development of mitochondrial transfer therapy for OP using iPSCs.
Funding Source: This work was supported by the National Research Foundation of Korea (NRF) and ministries including Science and ICT, Trade, Industry and Energy, Health & Welfare, and Food and Drug Safety (RS-2023-KH142779, RS-2019-NR040058).
