Directional differentiation of murine CD117+ hemopoietic stem cells into immature dendritic cells and their identification / 南方医科大学学报
Journal of Southern Medical University
; (12): 450-453, 2007.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-268108
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To establish a stable method for obtaining large quantity of highly purified immature dendritic cells (imDCs) in vitro, and identify the morphology, function and surface markers of the cells.</p><p><b>METHODS</b>CD117(+) hemopoietic stem cells (HSCs) were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor (SCF) and interleukin-3 (IL-3), the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL-10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry.</p><p><b>RESULTS</b>After 3, 5 and 7 days of culture in the presence of SCF+IL-3, the cells were expanded by 10.34-/+1.43, 22.65-/+2.71 and 54.39-/+3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-).</p><p><b>CONCLUSION</b>The method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.</p>
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Dendritic Cells
/
Hematopoietic Stem Cells
/
Microscopy, Electron, Scanning
/
Cell Differentiation
/
Cell Separation
/
Cells, Cultured
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Proto-Oncogene Proteins c-kit
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Cell Culture Techniques
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Cell Biology
/
Microscopy, Electron, Transmission
Type of study:
Prognostic study
Limits:
Animals
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2007
Document type:
Article