Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (6): 1061-1064.doi: 10.3969/j.issn.1673-8225.2011.06.024

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Methodological study on directional sensing and lateral pseudopodia formation during neutrophil polarity

Chen Hai-yang1, Cai Chun-qing1, Xie Cui-hua2, Luo Qiong1, Meng Xiao-jing1   

  1. 1Department of Occupational Health and Occupational Medicine, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou  510515, Guangdong Province, China
    2Department of Endocrinology and Metabolism, Nanfang Hospital, Guangzhou  510515, Guangdong Province, China
  • Received:2010-11-15 Revised:2010-12-30 Online:2011-02-05 Published:2011-02-05
  • Contact: Meng Xiao-jing, Doctor, Professor, Department of Occupational Health and Occupational Medicine, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, Guangdong Province, China zhumeng@fimmu.com
  • About author:Chen Hai-yang★, Master, Department of Occupational Health and Occupational Medicine, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, Guangdong Province, China ocean198511@sina.com Cai Chun-qing, Doctor, Associate professor, Department of Occupational Health and Occupational Medicine, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, Guangdong Province, China h_zcn@126.com Chen Hai-yang and Cai Chun-qing contributed equally to this paper.
  • Supported by:

    the National Natural Science Foundation of China, No. 30971193*, 30800438*, 81071611*

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Abstract:

BACKGROUND: Neutrophil chemotaxis is an important response in the host defense and initiated by neutrophil polarization. A key event of neutrophil polarity is the ability of directional sensing and lateral pseudopodium formation. However, there are few reports regarding the correlative investigation of the neutrophil polarization.
OBJECTIVE: To compare the three experimental modes of N-formyl-methionyl-leucyl-phenylalanine (Fmlp)-stimulated neutrophil polarity to provide some methodological considerations.
METHODS: ①Isolation of neutrophil by density gradient centrifugation; ②The neutrophil were stimulated by uniform concentration of fMLP, steep gradient concentration of fLMP and gentle gradient concentration of fLMP; ③The change and direction of neutrophil polarization were observed by inverted microscope; ④F-actin and cell nucleus were observed by confocal microscopy.
RESULTS AND CONCLUSION: ①The direction of neutronphil treated by uniform concentration of fMLP was random. ②41% of neutrophil migrated electrode-oriented when stimulated by steep gradient concentration of fLMP and 68% of neutrophil move toward the fMLP side upon stimulated by gentle gradient concentration of fLMP. ③χ2 test has been shown that there are significant difference in the polarized direction among neutrophils treated by three experimental modes. ④Neutronphil lateral pseudopodia formed in all of three experimental modes, especially in the group of uniform concentration. It is better to use zigmond chamber to study on the mechanism of neutrophils directional sensing, which could be interfered by the formation of neutrophil lateral pseudopodia.

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