Journal: Cell Death & Disease

Article Title: Mitochondrial transplantation: adaptive bio-enhancement

doi: 10.1038/s41419-025-07643-8

Figure Lengend Snippet: A Analysis of membrane potential in both cells and animals isolated mitochondria. Ctrl: Mitochondria were subjected to two freeze-thaw cycles at −80 °C and 37 °C to completely disrupt their structure and function, serving as a comparison, n = 3. B ATP production capacity in both cells and animals isolated mitochondria. Ctrl: Mitochondria were subjected to two freeze-thaw cycles at −80 °C and 37 °C, used for comparison, n = 3. C Mitochondria were randomly selected from cells (Vero) and animals (Bull Frog) for electron microscopy to observe their morphology. Scale bar: 500 nm. D – G Activity of respiratory chain complexes I and III in both cells and animals isolated mitochondria, n = 3. H Co-localization fluorescence imaging at 24 h of transplanted mitochondria labeled with Mito-Tracker Green (green fluorescence) from Eel, MDCK, CRFK, Sparrow, Vero, and Lizard, alongside AC16, L929, and HepG2 cells labeled with WGA594 (red fluorescence). Blue fluorescence (DAPI) represents the nucleus. Scale bar: 10 μm. I – N Immunoreactivity of IL-6, IL-10, and TNF-α in cell culture medium collected 24 h after the transplantation of exogenous mitochondria, n = 3. Ctrl: Untreated group. ns no statistical significance, **** p < 0.0001.

Article Snippet: Mitochondria were isolated from cultured cells, animal tissues, and plant tissues using the Mitochondria-Cytosol Protein Isolation Kit (Applygen) and the Plant Mitochondrial Extraction Kit (Solarbio).

Techniques: Membrane, Isolation, Comparison, Electron Microscopy, Activity Assay, Fluorescence, Imaging, Labeling, Cell Culture, Transplantation Assay

Journal: Cell Death & Disease

Article Title: Mitochondrial transplantation: adaptive bio-enhancement

doi: 10.1038/s41419-025-07643-8

Figure Lengend Snippet: A , I ATP production capacity in isolated mitochondria. Ctrl: Mitochondria subjected to two freeze-thaw cycles at −80 °C and 37 °C, used for comparison, n = 3. B , J Membrane potential in isolated mitochondria. Ctrl: Mitochondria subjected to two freeze-thaw cycles at −80 °C and 37 °C, used for comparison, n = 3. C Effects of mitochondrial transplantation from four different species on the viability of BMDM cells after H 2 O 2 treatment. Ctrl: Untreated group, n = 3. D Mitochondrial ROS fluorescence intensity in BMDM cells 24 h after transplantation of mitochondria from four different species, followed by H 2 O 2 treatment. Ctrl: Untreated group, n = 3. E , F Effects of mitochondrial transplantation from four different species on MDA and GSH levels in BMDM cells after H 2 O 2 treatment. Ctrl: Untreated group, n = 3. G , H Fluorescence imaging and statistical analysis of the internalization of mitochondria A labeled with Mito-Tracker Red and mitochondria B labeled with Mito-Tracker Green in BMDM cells after H 2 O 2 treatment. Original image scale bar: 10 µm; magnified region scale bar: 2 µm, n = 9. K Effects of mitochondrial transplantation from four different species on the viability of AC16 cells after CCCP treatment. Ctrl: Untreated group, n = 3. L – N Changes in IL-6, IL-10, and TNF-α levels in AC16 cells after CCCP treatment, following mitochondrial transplantation from four different species. Ctrl: Untreated group, n = 3. ns no statistical significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mitochondria were isolated from cultured cells, animal tissues, and plant tissues using the Mitochondria-Cytosol Protein Isolation Kit (Applygen) and the Plant Mitochondrial Extraction Kit (Solarbio).

Techniques: Isolation, Comparison, Membrane, Transplantation Assay, Fluorescence, Imaging, Labeling

Journal: Cell Death & Disease

Article Title: Mitochondrial transplantation: adaptive bio-enhancement

doi: 10.1038/s41419-025-07643-8

Figure Lengend Snippet: A Co-localization fluorescence imaging 24 h after the fusion of HL1 cells labeled with WGA647 and H9C2 cells labeled with WGA594. Scale bar: 20 µm. B Flow cytometric sorting of hybrid cells 24 h after the fusion of HL1 cells labeled with WGA647 and H9C2 cells labeled with WGA488. C , D Western blot analysis of the mitochondrial fusion and fission proteins Mfn1, Opa1, and Drp1 in HL1, H9C2, and HL1 + H9C2 cells on the second day after cell fusion, n = 3. E ATP production capacity in isolated mitochondria, Ctrl: Mitochondria subjected to two freeze-thaw cycles at −80 °C and 37 °C, used for comparison, n = 3. F JC-1 membrane potential detection of isolated mitochondria. Ctrl: Mitochondria subjected to two freeze-thaw cycles at −80 °C and 37 °C, used for comparison, n = 3. G Effects of transplantation of three different types of mitochondria on the viability of AC16 cells after H 2 O 2 treatment. Ctrl: Untreated group, n = 3. H Effects of transplantation of three different types of mitochondria on mitochondrial ROS levels in AC16 cells after H 2 O 2 treatment. Ctrl: Untreated group, n = 3. I , J Effects of transplantation of mitochondria from three different species on MDA and GSH levels in AC16 cells after H 2 O 2 treatment. Ctrl: Untreated group, n = 3. ns no statistical significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mitochondria were isolated from cultured cells, animal tissues, and plant tissues using the Mitochondria-Cytosol Protein Isolation Kit (Applygen) and the Plant Mitochondrial Extraction Kit (Solarbio).

Techniques: Fluorescence, Imaging, Labeling, Western Blot, Isolation, Comparison, Membrane, Transplantation Assay

Journal: Cell Death & Disease

Article Title: Mitochondrial transplantation: adaptive bio-enhancement

doi: 10.1038/s41419-025-07643-8

Figure Lengend Snippet: A , H ATP production capacity in isolated mitochondria. Ctrl: Mitochondria subjected to two freeze-thaw cycles at −80 °C and 37 °C, used for comparison, n = 3. B , I Membrane potential in isolated mitochondria. Ctrl: Mitochondria subjected to two freeze-thaw cycles at −80 °C and 37 °C, used for comparison, n = 3. C Effects of mitochondria transplantation from four different species on the viability of HepG2 cells after CCCP treatment. Ctrl: Untreated group, n = 3. D Effects of mitochondria transplantation from four different species on mitochondria ROS in HepG2 cells after CCCP treatment. Ctrl: Untreated group, n = 3. E Detection of DNA damage repair in HepG2 cells after CCCP treatment following mitochondria transplantation from four different species. Ctrl: Untreated group, n = 3. F , G Fluorescence imaging and statistical analysis of the internalization of Lizard mitochondria labeled with Mito-Tracker Deep-Red and Bull Frog mitochondria labeled with Mito-Tracker Green in HepG2 cells after CCCP treatment. Original image scale bar: 10 µm, magnified region scale bar: 2 µm, n = 6. J Effects of mitochondria transplantation from four different species on the viability of BMDM cells after LPS treatment. Ctrl: Untreated group, n = 3. K – M Changes in IL-6, IL-10, and TNF-α levels in BMDM cells after LPS treatment following mitochondria transplantation from four different species. Ctrl: Untreated group, n = 3. N Detection of DNA damage repair in BMDM cells after LPS treatment following mitochondria transplantation from four different species, n = 3. O , P Fluorescence imaging and statistical analysis of the internalization of Lizard mitochondria labeled with Mito-Tracker Green, Sparrow mitochondria labeled with Mito-Tracker Deep-Red, and Salmon mitochondria labeled with Mito-Tracker Red in BMDM cells after LPS treatment. Original image scale bar: 10 µm, magnified region scale bar: 5 µm, n = 6. ns no statistical significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mitochondria were isolated from cultured cells, animal tissues, and plant tissues using the Mitochondria-Cytosol Protein Isolation Kit (Applygen) and the Plant Mitochondrial Extraction Kit (Solarbio).

Techniques: Isolation, Comparison, Membrane, Transplantation Assay, Fluorescence, Imaging, Labeling

Journal: Cell Death & Disease

Article Title: Mitochondrial transplantation: adaptive bio-enhancement

doi: 10.1038/s41419-025-07643-8

Figure Lengend Snippet: A , B Evaluation of isolated mitochondrial functions through ATP production and mitochondrial membrane potential (MMP) assays, with mitochondria subjected to two freeze-thaw cycles at −80 °C and 37 °C to completely disrupt their structure and function as a control, n = 3. C Biodistribution of exogenous mitochondria in the mouse liver, kidney, and heart detected by in vivo imaging 24 h after intravenous tail vein injection of mitochondria. The blank group consists of untreated LPS-Ctrl group mice. D , E Assessment of motor function in acute inflammation model mice and normal model mice 3 days after the transplantation of two different types of mitochondria, with untreated groups as control, n = 6. F – H Changes in body weight of acute inflammation model mice and normal model mice before and 1 week after the transplantation of two different types of mitochondria, with untreated groups as control, n = 6. I Grip strength measurements in acute inflammation model mice and normal model mice 3 days after the transplantation of two different types of mitochondria, with untreated groups as control, n = 6. ns no statistical significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mitochondria were isolated from cultured cells, animal tissues, and plant tissues using the Mitochondria-Cytosol Protein Isolation Kit (Applygen) and the Plant Mitochondrial Extraction Kit (Solarbio).

Techniques: Isolation, Membrane, Control, In Vivo Imaging, Injection, Transplantation Assay

Journal: Cell Death & Disease

Article Title: Mitochondrial transplantation: adaptive bio-enhancement

doi: 10.1038/s41419-025-07643-8

Figure Lengend Snippet: A – C Changes in serum levels of IL-6, TNF-α, and IL-10 in acute inflammation model mice and normal model mice 1 week after transplantation of two different mitochondria, with untreated groups as control, n = 3. D – F Evaluation of kidney and liver function via serum Cr, ALT, and AST levels in acute inflammation model mice and normal model mice 1 week after transplantation of two different mitochondria, with untreated groups as control, n = 3. G , H Measurement of serum MDA levels and SOD activity in acute inflammation model mice and normal model mice 1 week after transplantation of two different mitochondria, with untreated groups as control, n = 3. I – K Analysis of JNK and P-JNK, p65, and P-p65 protein expression levels in liver and kidney tissues via Western blot 1 week after transplantation of two different mitochondria in acute inflammation model mice and normal model mice, with untreated groups as control, n = 3. L Effects of two different mitochondria on the viability of LPS-treated BMDM cells 24 h after transplantation, with untreated groups as control, n = 3. M Quantitative analysis of mitochondrial ROS levels in LPS-treated BMDM cells 24 h after transplantation of two different mitochondria, with untreated groups as control, n = 3. ns no statistical significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mitochondria were isolated from cultured cells, animal tissues, and plant tissues using the Mitochondria-Cytosol Protein Isolation Kit (Applygen) and the Plant Mitochondrial Extraction Kit (Solarbio).

Techniques: Transplantation Assay, Control, Activity Assay, Expressing, Western Blot

Journal: The Journal of Neuroscience

Article Title: Neuroprotection and Neuronal Differentiation Studies Using Substantia Nigra Dopaminergic Cells Derived from Transgenic Mouse Embryos

doi: 10.1523/JNEUROSCI.19-01-00010.1999

Figure Lengend Snippet: Detection of TH expression and DA neuronal markers by Western blot analysis and RT-PCR. A, The representative DA marker, TH expression, was confirmed by Western blot analysis after maintaining the SN4741 cells for a week in high-density culture at 37°C (lane 4) and 33°C (lane 5). The 62 kDa TH band in the SN4741 cells was consistent with the molecular weight of TH isolated from mouse adrenal gland (lane 1) and similarly derived locus coeruleus noradrenergic cell line, grown at 37°C (lane 2) and 33°C (lane 3), which were used as positive controls for the Western blot analysis. Each lane contained 15 μg of protein except lane 1 (5 μg) and lane 2 (30 μg). The specific amount of TH protein in the SN4741 cells grown at 37°C was higher than the culture at 33°C. As demonstrated in a TH-positive pheochromocytoma cell, PC12, culture (Kim et al., 1995), our SN DA cell line also exhibited the increase in TH expression in the high cell density during a prolonged culture (7–10 d). B, The expression of some DA neuron-specific markers, such as NT-3, BDNF, DA-T, and D2R were detected by RT-PCR in the SN4741 cell line. After Southern blotting of both 1 and 5 μl of each RT-PCR sample, the presence of each specific size band (the strongest band) was detected and confirmed with radiolabeled human BDNF cDNA, rat NT-3 cDNA, mouse DA-T cDNA, or mouse D2R oligonucleotide probe, respectively. Each filter was hybridized with each specific probe and exposed separately. The expected major band sizes were 430 bp for NT-3, 407 bp for BDNF, 553 bp for DA-T, and 490 bp for D2R. β represents the β-actin PCR product of 289 bp as an internal standard.

Article Snippet: PCR was performed with Taq polymerase (Perkin-Elmer, Norwalk, CT) using an appropriate primer sets for BDNF (nucleotides 374–393 and 761–780; GenBank accession number G287898), neurotrophin-3 (NT-3, nucleotides 441–460 and 851–870; GenBank accession number {"type":"entrez-nucleotide","attrs":{"text":"X53257","term_id":"53451"}} X53257 ), D 2 autoreceptor (D 2 R, nucleotides 1581–1600 and 2051–2070; GenBank accession number {"type":"entrez-nucleotide","attrs":{"text":"X55674","term_id":"50648"}} X55674 ), and dopamine transporter (DA-T, nucleotides 2381–2400 and 6241–6250; GenBank Accession number {"type":"entrez-nucleotide","attrs":{"text":"U15791","term_id":"338819863"}} U15791 ), a β-actin internal standard for normalization (Gene Link, Thornwood, NY), trace amounts of [ 32 P]dCTP, and various amounts of cDNA.

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Marker, Molecular Weight, Isolation, Derivative Assay, Southern Blot