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Efficient extraction of primary peritoneal macrophages in
mice induced by 3% thioglycollate broth
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GENG Tianxin, GENG Tian-xin, ZANG Guang-yao, YAN Jin-chuan |
(Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001, China)
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Abstract Objective: To establish an efficient method for the isolation of murine peritoneal macrophages by comparing the biological characteristics of mouse peritoneal macrophages with different methods. Methods: Thirtytwo healthy Kunming mice were selected and randomly divided into 4 groups, and macrophages were obtained by four different methods, i.e., PBS direct peritoneal lavage(PBS group), intraperitoneal injection of RPMI 1640 medium containing 10% serum for 30 min followed with PBS lavage(medium group), intraperitoneal injection of 6% starch broth for 3 d followed with PBS lavage(starch broth group) and intraperitoneal injection of 3% thioglycollate broth for 3 d followed with PBS lavage(thioglycollate group). The obtained cells were then transferred into the RPMI 1640 culture medium containing 10% serum and cultured. The cell morphology, number, phagocytic ability, purity and viability were compared. Results: There was no significant difference in the morphology of macrophages among the four groups; the cell number and the phagocytic ability in thioglycollate broth group were significantly higher than those of other three groups(t=16.685, 9.361, 3.199, all P0.05), which were greater than 95% respectively. Conclusion: Intraperitoneal injection of 3% thioglycollate broth for 3 d followed with PBS lavage, is a highly efficient method for obtaining mouse peritoneal macrophages.
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Received: 12 July 2018
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