Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (5): 854-857.doi: 10.3969/j.issn.1673-8225.2010.05.022

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In vitro amplification and ultrastructure of dendritic cells from mouse bone marrow

Wu Shu-yan1, Wang Xiang-ying2, Yang Gang2, Li Su-an2, Huang Rui1   

  1. 1 Medical School, 2 Center of Analysis and Measurement, Soochow University, Suzhou  215123, Jiangsu Province, China
  • Online:2010-01-29 Published:2010-01-29
  • Contact: Huang Rui, Doctor, Professor, Doctoral supervisor, Medical School, Soochow University, Suzhou 215123, Jiangsu Province, China hruisdm@163.com
  • About author:Wu Shu-yan☆, Doctor, Associate professor, Master’s supervisor, Medical School, Soochow University, Suzhou 215123, Jiangsu Province, China shuyanzw@sohu.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30972768* and General Specific Project of Chinese Postdoctoral Science Foundation, No. 2009025291*, 20080430178*; the Natural Science Foundation of Jiangsu High Education Institutes, No. 08KJB310009*; the Social Development Science Foundation of Suzhou City, No. SS08025*

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Abstract:

BACKGROUND: Dendritic cells (DCs) constitute the dominant population of antigen presenting cells (APCs) by possessing potent ability to initiate T cell immunity. The ultrastructure study of DCs is less reported.
OBJECTIVE: To investigate the ultrastructure of DCs from mice bone marrow at different maturation stages, and the morphology of DCs between CD40 ligation and tumor necrosis factor-alpha (TNF-α) stimulation in vitro.
METHODS: Mice myeloid DCs were generated from bone marrow in vitro using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Immature DCs were loaded with apoptotic tumor cells (AP-DC), and AP-DC was then stimulated with CD40L-CHO cells and TNF-α for 48 hours, respectively. DCs were routinely sectioned, and ultrastructure was observed under transmission electron microscope.
RESULTS AND CONCLUSION: Immature DCs showed a few short and blunt cytoplasmic processes, there were specific morphology lysosomes that liked earphone in some cells; DCs engulfing the apoptotic bodies were observed; sub-cellular structures between CD40 ligation and TNF-α stimulated DCs were different, the former had typical morphology of mature DCs which exhibited many dendritic protrusions, however, some DCs displayed apoptosis and autophagy after TNF-α stimulation. In a conclusion, CD40 ligation plays an essential role in myeloid DCs differentiation and maturation, TNF-α can mediate apoptosis and autophagy of DCs.

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