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BALB/C小鼠骨髓源性树突状细胞的 定向诱导分化及鉴定.
- Source :
-
Journal of Xi'an Jiaotong University (Medical Sciences) . Mar2021, Vol. 42 Issue 2, p219-224. 6p. - Publication Year :
- 2021
-
Abstract
- To establish an effective method for acquiring bone marrow-derived dendritic cells (DCs) from BALB/C mice in vitro and to establish a reservoir of DC precursor cells. Methods CD117+ hematopoietic stem cells (HSCs) were isolated and purified from bone marrow of BALB/C mice by immunomagnetic beads separation system (MACS), and then amplified in vitro with mouse stem cell factor (SCF) and interleukin-3 (IL-3). HSC was induced to differentiate into DCs by adding granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and IL-4. Different cytokines (tumor necrosis factor-alpha or IL-10) were added to control the maturity of dendritic cells. Then the morphology (electron microscopy), surface molecular markers (FACS method) and cytokine secretion level (ELISA method) were identified. Results ① The purity of CD117 + HSC isolated and purified by MACS system was over 95%. ② SCF plus IL-3 could effectively stimulate HSC amplification. ③ The morphology of mature DC (mDC) and immature DC (imDC) was significantly different under light and scanning electron microscopy. ④ In the expressions of surface markers CD40, CD80, CD86, I-A/I-E, there were significant differences between imDC group and mDC group (P<0.01). ⑤ After LPS stimulation, the secretion of IL-12 in imDC group did not change significantly (P=0.064), while the secretion of IL-12 in mDC group increased significantly (P=0.009). LPS and TNF-α had a synergistic effect in stimulating DC maturation. Conclusion Specific combinations of cytokines can effectively induce the differentiation of bone marrow HSCs into DCs in BALB/C mice, and can control the maturity of DCs. This study makes it possible to use gene modified dendritic cells in GD immunotherapy. [ABSTRACT FROM AUTHOR]
Details
- Language :
- Chinese
- ISSN :
- 16718259
- Volume :
- 42
- Issue :
- 2
- Database :
- Academic Search Index
- Journal :
- Journal of Xi'an Jiaotong University (Medical Sciences)
- Publication Type :
- Academic Journal
- Accession number :
- 150239240
- Full Text :
- https://doi.org/10.7652/jdyxb202102009