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《China Biotechnology》 2015-01
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c-Kit~+ Combining with DiI Micro-label to Sort the Cardiac Telocytes

XIAO Hu-song;SHEN Xiao-tao;ZHENG Xin;LI Yan-mei;CAI Dong-qing;Key Laboratory for Regenerative Medicine,Ministry of Education,Jinan University;The International Base of Collaboration for Science & Technology,The Ministry of Science and Technology & Guangdong Province,Jinan University;Department of Developmental and Regenerative Biology,Ji Nan University;  
Objective: To establish methodology for specific sorting of cardiac telocytes using c-Kit+Combining with Di I Micro-label and flow cytometry. Methods: The cardiac telocytes were isolated via a c-Kit antibody conjugated magnetic bead method. The cells which have unique telocyte morphology, with long"Telopode",were labeled with Di I microinjection. The labeling Di I+cells were sorted and recovered in 96-well plate by single cell flow cytometry technique. The immunofluorescence and RT-PCR were applied to confirm the phenotype of collected cells. Results: The cardiac telocytes( c-Kit+) which have unique telocyte morphology were able to be labeled by Di I microinjection. The Di I labeled cells were able to be sorted and recovered in single cell level. The adherent cells of collected Di I+cells were 14. 9% under culture. The 5. 6% of cultured Di I+cells was able to proliferate and the 2. 4% of cultured Di I+cells was able to form cell-clone. In addition,the formed cell-clone was able to subculture. The immunofluorescence staining confirmed that the recovered Di I+cells were c-Kit-and CD34-positive. RT-PCR results also showed that the recovered Di I+cells were positive for cKit,CD34,Vimentin and PDGFR-β,which are the markers for telocytes. Conclusion: The methodology which was established in present study was able to isolate and sort cardiac telocytes in single cell level. The sorted cardiac telocytes were able to proliferate and maintain the unique phenotype.
【Fund】: 科技部港澳台合作专项(2012DFH30060);; 国家自然科学基金(81170324 81470433);; 广东省教育厅国际合作平台项目(2012gjhz0003);; 广东省自然科学基金重点项目(S2012020010895);; 广东省科技厅国际合作平台项目(2013B051000062)资助项目
【CateGory Index】: R329.2
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