Mitochondria in Transfer: Intercellular Delivery Driving Tumor Progression [Sep 30, 2025] DOJINDO LABORATORIES

Previous Science Note

Recent studies have drawn attention to intercellular mitochondrial transfer in cancer. Tumors have been reported to promote intratumoral innervation through neurotrophic factors, and recent work shows that mitochondria supplied by neurons shift cancer cell metabolism and are linked to increased metastatic potential. Moreover, for cancer-associated fibroblasts (CAFs), known supporters of tumor progression, transfer of mitochondria from cancer cells has been shown to induce CAF differentiation, deepening our understanding of intercellular mitochondrial transfer as a strategy in cancer.

Nerve-to-cancer transfer of mitochondria during cancer metastasis (Nature, 2025)
Summary: This study revealed that nerves within tumors transfer their mitochondria to cancer cells, enhancing their metabolic flexibility and driving more aggressive, metastatic behavior. By permanently tracing this event, the researchers showed that cancer cells receiving neuronal mitochondria were enriched at metastatic sites and linked to increased malignancy.

Highlighted technique: MitoTRACER is a genetic switch built into cancer cells, containing a reporter with DsRed and eGFP. The cells normally fluoresce red, but when they receive mitochondria carrying Cre recombinase from neurons, Cre edits their DNA to switch fluorescence from red to green. This irreversible change is inherited by daughter cells, enabling permanent lineage tracing of cancer cells that acquired neuronal mitochondria.

Related technique Mitochondria Detection, Glycolysis/OXPHOS Assay

MIRO2-mediated mitochondrial transfer from cancer cells induces cancer-associated fibroblast differentiation (Nature Cancer, 2025)
Summary: Cancer-associated fibroblasts (CAFs) are well-known supporters of tumor growth, but how cancer cells convert normal fibroblasts into CAFs has remained unclear. This study demonstrates that cancer cells transfer mitochondria to fibroblasts in a process requiring the trafficking protein MIRO2, reprogramming them into protumorigenic CAFs that promote tumor progression.

Highlighted technique: The authors isolated mitochondria from cancer cells and transplanted them into fibroblasts, directly testing whether mitochondria alone could reprogram fibroblasts into CAF-like cells. Using Seahorse assays, they showed that the transplanted mitochondria increased oxidative phosphorylation (OCR) and ATP production, confirming a functional metabolic shift in recipient fibroblasts.

Related technique OCR Plate Assay, ATP Assay

Related Techniques (click to open/close)

Target	Kit & Probes
Mitochondrial Staining	MitoBright LT Green / Red / Deep Red
Mitochondrial membrane potential detection	JC-1 MitoMP Detection Kit, MT-1 MitoMP Detection Kit
Mitochondrial superoxide detection	MitoBright ROS Deep Red - Mitochondrial Superoxide Detection
Mitophagy detection	Mitophagy Detection Kit
Total ROS detection	Highly sensitive DCFH-DA or Photo-oxidation Resistant DCFH-DA
Glycolysis/Oxidative phosphorylation Assay	Glycolysis/OXPHOS Assay Kit
Oxygen consumption rate assay	Extracellular OCR Plate Assay Kit
Intracellular ATP Assay	ATP Assay Kit-Luminescence
Cell proliferation/ cytotoxicity assay	Cell Counting Kit-8 and Cytotoxicity LDH Assay Kit-WST

Application Note I (click to open/close)
> Inhibition of Mitochondrial Electron Transport Chain

Antimycin stimulation of Jurkat cells was used to evaluate the changes in cellular state upon inhibition of the mitochondrial electron transport chain using a variety of indicators.

The results showed that inhibition of the electron transport chain resulted in (1) a decrease in mitochondrial membrane potential and (2) a decrease in OCR. In addition, (3) the NAD⁺/NADH ratio of the entire glycolytic pathway decreased due to increased metabolism of pyruvate to lactate to maintain the glycolytic pathway, (4) GSH depletion due to increased reactive oxygen species (ROS), and (6) increase in the NADP⁺/NADPH ratio due to decreased NADH required for glutathione biosynthesis were observed.

Application Note II (click to open/close)
> Tracking ROS and Membrane Potential Decline

After HeLa cells were washed with HBSS, co-stained with MitoBright ROS Deep Red and mitochondrial membrane potential staining dye (JC-1: code MT09), and the generated mitochondrial ROS and membrane potential were observed simultaneously. As a result, the decrease in mitochondrial membrane potential and the generation of mitochondrial ROS are simultaneously observed.

＜LEFT: Imaging Conditions＞(Confocal microscopy)
JC-1: Green Ex = 488, Em = 490-520 nm, Red: Ex = 561, Em = 560-600 nm
MitoBright ROS ：Ex = 633 nm, Em = 640-700 nm
Scale bar: 10 μm

＜Right: Examination Conditions＞(Plate Reader)Tecan, Infinite M200 Pro
JC-1: Green Ex=480-490 nm, Em=525-545 nm; Red: Ex= 530-540 nm, Em=585-605 nm
MitoBright ROS: Ex=545-555 nm, Em = 665-685 nm