[Immature dendritic cells directional differentiated from murine hemopoietic stem cells and morphous and function of them by MACS].
OBJECTIVE
To Investigate a method that can obtain massive highly purified immature dendritic cells(imDCs) steadily in vitro, and identify them by morphous, function and surface markers by MACS.


METHODS
Isolate and purify CD117(+) hemopoietic stem cells(HSCs) from bone marrow of healthy C57 murine by MACS. After being expanded by SCF+IL-3, HSCs would be directional differentiated into imDCs by use of cytokine scheme of GM-CSF+IL-4+IL-10. Then identify imDCs through following ways: observing morphous and function of them under inverted microscope, scanning electron microscope and transmission electron microscope, detecting the expression of surface markers by flow cytometry.


RESULTS
The fold of expansion 3, 5 and 7 days after cultured with SCF+IL-3 was separately 10.34 +/- 1.43, 22.65 +/- 2.71 and 54.39 +/- 3.08. HSCs can be successfully differentiated into imDCs, which have the function of phagocytosis. The imDCs were short and small, in shape of sentus. The expression of surfacee markers was CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-) by flow cytometry.


CONCLUSIONS
This method can obtain and identify massive highly purified imDCs steadily in vitro.
Jiang
Ya-Zhuo
YZ
Department of Renal Transplantation, the First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, China. jerry_jyz@yahoo.com.cn
Tian
Pu-Xun
PX
Ding
Xiao-Ming
XM
Xue
Wu-Jun
WJ
Ding
Chen-Guang
CG
D000818:Animals; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D008280:Magnetics; D008297:Male; D051379:Mice; D008855:Microscopy, Electron, Scanning; D010587:Phagocytosis; D013234:Stem Cells / Q000166:cytology*
D015415:Biomarkers