Dongli Song1, Menglin Xu1, Ruixue Qi1, Ruihua Ma1, Yile Zhou1, Duojiao Wu2,3, Hao Fang4,5, Xiangdong Wang6,7. 1. Zhongshan Hospital Institute of Clinical Science, Shanghai Institute of Clinical Bioinformatics, Shanghai Engineering Research for AI Technology for Cardiopulmonary Diseases, Shanghai, China. 2. Zhongshan Hospital Institute of Clinical Science, Shanghai Institute of Clinical Bioinformatics, Shanghai Engineering Research for AI Technology for Cardiopulmonary Diseases, Shanghai, China. wuduojiao@126.com. 3. Center for Tumor Diagnosis and Therapy, Zhongshan Hospital Jinshan Hospital, Shanghai Medical College, Fudan University, Shanghai, China. wuduojiao@126.com. 4. Department of Anesthesiology, Zhongshan Hospital, Shanghai, China. drfanghao@163.com. 5. Department of Anesthesiology, Minhang Branch, Zhongshan Hospital, Fudan University, Shanghai, China. drfanghao@163.com. 6. Zhongshan Hospital Institute of Clinical Science, Shanghai Institute of Clinical Bioinformatics, Shanghai Engineering Research for AI Technology for Cardiopulmonary Diseases, Shanghai, China. xdwang@fuccb.com. 7. Center for Tumor Diagnosis and Therapy, Zhongshan Hospital Jinshan Hospital, Shanghai Medical College, Fudan University, Shanghai, China. xdwang@fuccb.com.
Abstract
BACKGROUND: Telocytes play key roles in maintenance of organ/tissue function and prevention of organ injury. However, there are great challenges to investigate telocytes functions using primary telocytes, due to the difficulties of isolation, identification, and stability. The present study aims at constructing continuous cell strain of mouse lung telocyte cell line with stable characters by gene modification and investigating biological behaviors and responses of gene-modified telocytes to inflammation. METHODS: Mouse primary lung telocytes were isolated and identified using immune-labeling markers and immunoelectron microscopy. Primary telocytes were transformed with Simian vacuolating virus 40 small and large T antigen (SV40). Biological characters, behaviors morphology, and proliferation of those gene-modified telocytes were defined and monitored dynamically for 50 generations, as compared with primary lung telocytes. Cell cycle of mouse primary lung telocytes or gene-modified telocytes was detected by flow cytometry. RESULTS: Gene modified telocytes of generations 5, 10, 30 and 50 were observed with telopodes and also showed CD34 and ckit positive. Multiple cellular morphology were also observed on telocyte cell-line under monitor of celliq and enhanced cell proliferation were showed. SV40 transduction was also reduced apoptosis and increased the ratio of S and G2 phases in telocyte cell-line. CONCLUSION: We successfully constructed mouse lung telocyte cell-line which maintained the biological properties and behaviors as primary telocytes and could responses to inflammation induced by LPS. Thus, gene-modified lung telocytes, Telocyte Line, would provide a cell tool for researchers exploring the roles and applications of telocytes involved in physiological and pathological states in future.
BACKGROUND: Telocytes play key roles in maintenance of organ/tissue function and prevention of organ injury. However, there are great challenges to investigate telocytes functions using primary telocytes, due to the difficulties of isolation, identification, and stability. The present study aims at constructing continuous cell strain of mouse lung telocyte cell line with stable characters by gene modification and investigating biological behaviors and responses of gene-modified telocytes to inflammation. METHODS:Mouse primary lung telocytes were isolated and identified using immune-labeling markers and immunoelectron microscopy. Primary telocytes were transformed with Simian vacuolating virus 40 small and large T antigen (SV40). Biological characters, behaviors morphology, and proliferation of those gene-modified telocytes were defined and monitored dynamically for 50 generations, as compared with primary lung telocytes. Cell cycle of mouse primary lung telocytes or gene-modified telocytes was detected by flow cytometry. RESULTS: Gene modified telocytes of generations 5, 10, 30 and 50 were observed with telopodes and also showed CD34 and ckit positive. Multiple cellular morphology were also observed on telocyte cell-line under monitor of celliq and enhanced cell proliferation were showed. SV40 transduction was also reduced apoptosis and increased the ratio of S and G2 phases in telocyte cell-line. CONCLUSION: We successfully constructed mouse lung telocyte cell-line which maintained the biological properties and behaviors as primary telocytes and could responses to inflammation induced by LPS. Thus, gene-modified lung telocytes, Telocyte Line, would provide a cell tool for researchers exploring the roles and applications of telocytes involved in physiological and pathological states in future.
Authors: Miguel G Toscano; Jeroen van der Velden; Sybrand van der Werf; Machteld Odijk; Ana Roque; Rafael J Camacho-Garcia; Irene G Herrera-Gomez; Irene Mancini; Peter de Haan Journal: Mol Ther Methods Clin Dev Date: 2017-07-05 Impact factor: 6.698
Authors: Payam A Gammage; Carlo Viscomi; Marie-Lune Simard; Ana S H Costa; Edoardo Gaude; Christopher A Powell; Lindsey Van Haute; Beverly J McCann; Pedro Rebelo-Guiomar; Raffaele Cerutti; Lei Zhang; Edward J Rebar; Massimo Zeviani; Christian Frezza; James B Stewart; Michal Minczuk Journal: Nat Med Date: 2018-09-24 Impact factor: 53.440