Mitochondria Dynamically Transplant into Cells in Vitro and in Mice and Rescue Aerobic Respiration of Mitochondrial DNA-Depleted Motor Neuron NSC-34  

作　　者：Xian-Peng Jiang Catherine C. Baucom Robert L. Elliott Xian-Peng Jiang;Catherine C. Baucom;Robert L. Elliott(Sallie A. Burdine Breast Foundation, Baton Rouge, LA, USA;Elliott Mastology Center, Baton Rouge, LA, USA)

机构地区：[1]Sallie A. Burdine Breast Foundation, Baton Rouge, LA, USA [2]Elliott Mastology Center, Baton Rouge, LA, USA

出　　处：《Journal of Biomedical Science and Engineering》2020年第9期203-221,共19页生物医学工程（英文）

摘　　要：It has been reported that transplantation of pheochromocytoma P12 and hepatoma cells’ mitochondria improve the locomotive activity and prevent disease progress in experimental Parkinson’s disease rats. To prepare for mitochondrial transplantation study in human neurodegenerative diseases, we select human fibroblasts as mitochondrial donor because that fibroblasts share many characteristics with mesenchymal stromal cells (MSCs). We isolate human primary fibroblasts and develop a mitochondrial DNA (mtDNA)-depleted mouse motor neuron NSC-34 cells (NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells). Fibroblast and NSC-34 cell’s mitochondria are co-cultured with NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells. Mitochondrial transplantation is observed by fluorescent microscopy. Gene expression is determined by polymerase chain reaction (PCR) and real time PCR (qPCR). Also, mitochondria are injected to mice bearing mammary adenocarcinoma 4T1 cells. We find results as following: 1) There are abundant mitochondria in fibroblasts (337 ± 80 mitochondria per fibroblast). 42.4% of viable mitochondria are obtained by using differential centrifugation. The isolated mitochondria actively transplant into NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells after co-culture. 2) Fibroblasts transfer mitochondria to human mammary adenocarcinoma MCF-7 cells. 3) There is no expression of HLA-I antigen in fibroblast’s mitochondria indicating they can be used for allogeneic mitochondrial transplantation without HLA antigen match. 4) PCR and qPCR show that NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells lose mitochondrially encoded cytochrome c oxidase I (MT-CO1) and mitochondrially encoded NADH dehydrogenase 1 (MT-ND1) and upregulate expression of glycolysis-associated genes hexokinase (HK2), glucose transporter 1 (SLC2A1) and lactate dehydrogenase A (LDHA). 5) Transplantation of NSC-34 mitochondria restores MT-CO1 and MT-ND1 and downregulIt has been reported that transplantation of pheochromocytoma P12 and hepatoma cells’ mitochondria improve the locomotive activity and prevent disease progress in experimental Parkinson’s disease rats. To prepare for mitochondrial transplantation study in human neurodegenerative diseases, we select human fibroblasts as mitochondrial donor because that fibroblasts share many characteristics with mesenchymal stromal cells (MSCs). We isolate human primary fibroblasts and develop a mitochondrial DNA (mtDNA)-depleted mouse motor neuron NSC-34 cells (NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells). Fibroblast and NSC-34 cell’s mitochondria are co-cultured with NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells. Mitochondrial transplantation is observed by fluorescent microscopy. Gene expression is determined by polymerase chain reaction (PCR) and real time PCR (qPCR). Also, mitochondria are injected to mice bearing mammary adenocarcinoma 4T1 cells. We find results as following: 1) There are abundant mitochondria in fibroblasts (337 ± 80 mitochondria per fibroblast). 42.4% of viable mitochondria are obtained by using differential centrifugation. The isolated mitochondria actively transplant into NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells after co-culture. 2) Fibroblasts transfer mitochondria to human mammary adenocarcinoma MCF-7 cells. 3) There is no expression of HLA-I antigen in fibroblast’s mitochondria indicating they can be used for allogeneic mitochondrial transplantation without HLA antigen match. 4) PCR and qPCR show that NSC-34 <em>ρ</em><span style="white-space:nowrap;">&#176;</span> cells lose mitochondrially encoded cytochrome c oxidase I (MT-CO1) and mitochondrially encoded NADH dehydrogenase 1 (MT-ND1) and upregulate expression of glycolysis-associated genes hexokinase (HK2), glucose transporter 1 (SLC2A1) and lactate dehydrogenase A (LDHA). 5) Transplantation of NSC-34 mitochondria restores MT-CO1 and MT-ND1 and downregul

关 键 词：Mitochondrial Transplantation Motor Neuron MITOCHONDRIA Neurodegenerative Disease Mammary Adenocarcinoma Mitochondrial DNA Depletion Fibroblast HLA-I NSC-34 Cells 

分 类 号：R73[医药卫生—肿瘤]