Directional differentiation and identification of bone marrow-derived dendritic cells in BALB/C mice

Wenxu Zhou; Xiaoyan WU; Bingyin Shi; Ning NI; Chenguang Ding; Zhihui Guan; Yazhuo Jiang; Heli Xiang

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Directional differentiation and identification of bone marrow-derived dendritic cells in BALB/C mice

VernacularTitle:BALB/C小鼠骨髓源性树突状细胞的定向诱导分化及鉴定
Author: Wenxu ZHOU ^{1
,

2} ; Xiaoyan WU ¹ ; Bingyin SHI ¹ ; Ning NI ³ ; Chenguang DING ¹ ; Zhihui GUAN ¹ ; Yazhuo JIANG ¹ ; Heli XIANG ¹
Author Information

1. Department of Endocrinology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061
2. Department of Internal Medicine, The Hospital of Xi’an Jiaotong University, Xi’an 710049, China
3. Department of Internal Medicine, The Hospital of Xi’an Jiaotong University, Xi’an 710049, China
Publication Type:Journal Article
Keywords: BALB/C mouse; CD117⁺ hematopoietic stem cell; dendritic cell; directional differentiation
From: Journal of Xi'an Jiaotong University(Medical Sciences) 2021;42(2):219-224
CountryChina
Language:Chinese
Abstract: 【Objective】 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.