On March 17, 2025, Professor Ouyang Hongwei of Zhejiang University/Liangzhu Laboratory published a paper titled "Organelle-tuning condition robustly fabricates energetic mitochondria for Bone Research. In the research paper "cartilage regeneration", the research team built an exclusive "stem cell factory" for mitochondrial production by preparing an "cartilage regeneration" culture plan, which achieved high throughput and high-quality human mitochondria manufacturing, providing a new solution for the treatment of osteoarthritis, a degenerative disease.

Mitochondria are the "power stations" of the cell, responsible for providing 90% of the body's biological energy every day. Once the mitochondria is damaged or abnormal, all kinds of metabolic and degenerative diseases will take advantage of the opportunity to enter, seriously threatening human health. Mitochondrial transfer therapy is designed to deliver healthy mitochondria to diseased tissue, or to replace damaged mitochondria, or to activate cellular repair programs.

Studies have shown that mitochondrial transplantation has been or will be widely used in the treatment of many diseases, including ischemic heart disease, mitochondrial disease and so on. However, this therapy suffers from the "raw material" dilemma: each patient needs 109 levels of mitochondria per injection, and the current way to obtain mitochondria from unamplified somatic cells is not sustainable, mitochondrial quality is unstable, and is greatly affected by the age and health of the donor. Therefore, it is urgent to develop sustainable, controlled strategies for high-quality mitochondrial manufacturing.

"Mitochondrial factories" enable tissue regeneration

Building a "stem cell factory" for mitochondria manufacturing

In order to construct mitochondria manufacturing platform, the research team screened mesenchymal stem cells (MSCs) culture conditions to find components that can promote cell proliferation and mitochondrial quantity/activity, and defined their combination formula as mitochondria-producing condition. MSCs (mc-MSCs) cultured under this condition had significantly higher cell proliferation rate and mitochondrial number than MSCs (tc-MSCs) cultured with traditional serum.

Customized culture conditions promote mitochondrial biosynthesis

Sustainable production and functional enhancement of mitochondria

Based on the self-renewing properties of stem cells, the research team found that mitochondria can be produced steadily and sustainably in mc-MSCs. In about 15 days (5 times of passage), the number of cells obtained by mc-MSCs was 302 times that of traditional culture conditions, and the mitochondrial production was estimated to be 854 times that of traditional methods!

Sustainable mitochondrial production

In addition to the increase in number, mc-Mitochondria produced under optimized conditions showed higher mitochondrial activity (including ATP production level and mitochondrial membrane potential) both inside and outside the cells.

Adequate mitochondrial energy to ensure the transplantation effect

Activate the productivity pathway and fine-tune the organelle balance

In terms of mechanism, the research team found through transcriptome sequencing that mc-MSCs are enriched in genes related to mitochondrial biogenesis, cell proliferation and metabolism. The AMPK pathway, which is highly related to energy perception, is activated, and its downstream key transcription factor TFAM is up-regulated, promoting mitochondrial replication and transcription.

mito-condition activates AMPK pathway to promote mitochondrial biosynthesis

While activating AMPK pathway, mito-condition also altered the subcellular components and metabolic levels of mc-MSCs. The data show that mc-MSCs "intelligently" invest a large amount of intracellular resources into the cell cycle, mitochondrial synthesis, and energy production processes, while the remaining energy-consuming cellular behaviors unrelated to mitochondrial production, such as lysosomal function, vesicular secretion behavior, cell adhesion, and migration, are weakened. Even when autophagy levels are reduced and organelles are redistributed by constant cell division, mc-MSCs can maintain high energy homeostasis and avoid overloading of single organelles or metabolic waste. This new balance makes mc-MSCs a "celestial mitochondrial factory."

Restrain energy-consuming process and regulate energy balance

Building a "stem cell factory" for mitochondria manufacturing

Finally, the research team chose osteoarthritis as a therapeutic application scenario. Osteoarthritis is a serious, disabling disease that affects more than 500 million people worldwide. Studies have shown that it is related to the imbalance of chondrocyte metabolism, and mitochondrial transplantation is expected to promote cartilage regeneration and prevent osteoarthritis. First, in the mitochondrial production phase, mito-condition can produce the mitochondria needed for in vivo experiments with less time and money. Moreover, in mouse osteoarthritis model, mc-Mitochondria perform better than common mitochondria in maintaining cartilage phenotype, reducing inflammation and alleviating subchondral osteosclerosis, etc., even with the same number of injections, fully proving their value in the treatment of diseases in vivo.

High energy mitochondrial transplantation has excellent efficacy in vivo

In conclusion, this study establishes an efficient mitochondrial production platform, which can meet the needs of clinical treatment and provide innovative strategies for the prevention and treatment of degenerative diseases in the context of global aging. In addition, the study proposes a new paradigm of "organelle regulation", which provides new insights into the molecular regulation of intracellular matter and energy balance, and is expected to provide references for the development of new organelle therapy methods. In the future, single-cell spatiotemporal omics, high-definition imaging and other technologies will be further used to analyze the regulation law, and promote the application of organelle therapy in aging intervention and regenerative medicine.