Establishment of a serum-free primary culture method for enteric glial cell of myenteric plexus

doi:10.3969/j.issn.1006-7795.2019.02.015

Journal of Capital Medical University ›› 2019, Vol. 40 ›› Issue (2): 232-236.doi: 10.3969/j.issn.1006-7795.2019.02.015

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Establishment of a serum-free primary culture method for enteric glial cell of myenteric plexus

Quan Zhusheng, Zhang Xiaoli, Zhu Jinxia

Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China

Received:2018-11-27 Online:2019-03-21 Published:2019-04-15
Supported by:
This study was supported by The National Key Research and Development Program of China(2016YFC1302203), National Natural Science Foundation of China (81700462,31871159,81570695).

Abstract

Abstract: Objective Enteric glial cell (EGC) plays an important role in regulation of many physiological and pathological process in the intestine, in order to further study the function of EGC and exclude the influence of neurohumoral factors at the vivo and vitro level, it is necessary to obtain a simple and efficient method for isolation and primary culture of EGC. Methods Colonic myenteric plexus were digested and separated. The fibroblasts were removed by alternating culture with serum-containing and serum-free medium and then subcultured routinely. The purity and activity of primary EGC were detected by immunofluorescence and intracellular Ca²⁺ imaging. Results The number of EGC was about 6.8×10⁵ on each experiment before passage. After serum-free culture, fibroblasts decreased significantly; the percentage of immunoreactivities of glial fibrillary acidic protein(GFAP), S100β and both GFAP and S100β was 98.44%±1.07%, 93.61%±3.16% and 93.09%±2.99%, respectively.About 96% EGC manifested intracellular calcium transient when treated with ATP. Conclusion Combined with the serum-free culture,fibroblasts were removed and primary myenteric EGC with higher purity and activity could be obtained.

Key words: enteric glial cell, primary culture, serum-free culture

CLC Number:

R333.3

Quan Zhusheng, Zhang Xiaoli, Zhu Jinxia. Establishment of a serum-free primary culture method for enteric glial cell of myenteric plexus[J]. Journal of Capital Medical University, 2019, 40(2): 232-236.

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Establishment of a serum-free primary culture method for enteric glial cell of myenteric plexus

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