c-Kit<sup>+</sup> Combining with DiI Micro-label to Sort the Cardiac Telocytes

doi:10.13523/j.cb.20150107

China Biotechnology

2015, Vol. 35

Issue (1): 46-53 DOI: 10.13523/j.cb.20150107

c-Kit⁺ Combining with DiI Micro-label to Sort the Cardiac Telocytes

XIAO Hu-song^1,2,3, SHEN Xiao-tao^1,2,3, ZHENG Xin^1,2,3, LI Yan-mei^1,2,3, CAI Dong-qing^1,2,3

1. Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou 510632, China;
2. The International Base of Collaboration for Science & Technology, The Ministry of Science and Technology & Guangdong Province, Jinan University, Guangzhou 510632, China;
3. Department of Developmental and Regenerative Biology, Ji Nan University, Guangzhou 510632, China

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Abstract

Objective: To establish methodology for specific sorting of cardiac telocytes using c-Kit⁺ Combining with DiI 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 DiI microinjection. The labeling DiI⁺ 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 DiI microinjection. The DiI labeled cells were able to be sorted and recovered in single cell level. The adherent cells of collected DiI⁺ cells were 14.9% under culture. The 5.6% of cultured DiI⁺ cells was able to proliferate and the 2.4% of cultured DiI⁺ cells was able to form cell-clone. In addition, the formed cell-clone was able to subculture. The immunofluorescence staining confirmed that the recovered DiI⁺ cells were c-Kit-and CD34-positive. RT-PCR results also showed that the recovered DiI⁺ cells were positive for c-Kit, 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.

Key words： Cardiac telocytes Microinjection Single cell sorting

Received: 22 October 2014 Published: 25 January 2015

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XIAO Hu-song, SHEN Xiao-tao, ZHENG Xin, LI Yan-mei, CAI Dong-qing. c-Kit⁺ Combining with DiI Micro-label to Sort the Cardiac Telocytes. China Biotechnology, 2015, 35(1): 46-53.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150107 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I1/46

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