Int J Stomatol

Int J Stomatol ›› 2020, Vol. 47 ›› Issue (3): 286-292.doi: 10.7518/gjkq.2020062

• Original Articles • Previous Articles     Next Articles

Comparison of polarization characteristics of human monocyte cell- and peripheral blood mononuclear cell-derived macrophages

Liu Ye1,2,Hong Rundan2,Wang Zhiguo3,Liu Hanyun4,Meng Chenda2,Wang Ru2,Xu Quanchen1()   

  1. 1.Dept. of Oral Medicine, The Affiliated Hospital of Qingdao University, Qingdao 266003, China;
    2.School of Stomatology, Qingdao University, Qingdao 266003, China;
    3.Dept. of Burn and Plastic Surgery, The Affiliated Hospital of Qingdao University, Qingdao 266003, China;
    4.Dept. of Infectious Diseases, The Affiliated Hospital of Qingdao University, Qingdao 266003, China
  • Received:2019-09-17 Revised:2019-11-22 Online:2020-05-01 Published:2020-05-08
  • Contact: Quanchen Xu E-mail:qyfyxqc@126.com
  • Supported by:
    National Natural Science Foundation of China(81500849);Shandong Province Key Research Plan(2018GSF118150);Shandong Provincial Natural Science Foundation(ZR2017MH083)

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Abstract:

Objective Obtained from successfully established polarization models, the polarization characteristics of macrophages (Mφ) from different sources were compared to provide an experimental basis for biological treatments. Methods Peripheral blood mononuclear cells (PBMC) were isolated through density gradient centrifugation and magnetic cell sorting and were cultured in vitro. PBMC Mφ was induced by human recombinant macrophage colony-stimulating factor. The human monocyte cell line THP-1 was differentiated into THP-1 Mφ under the induction of 100 nmol·L-1 phorbol 12-myristate 13-acetate. Morphology, polarization marker expression (CD86 and CD206), and cytokine secretion levels of the two groups were compared under the condition of M1 or M2 polarization factors. M1 polarization factors was 50 ng·mL-1 lipopolysaccharides and 20 ng·mL-1 interferon-γ, and M2 polarization factors was 20 ng·mL-1 interleukin-4. Results After M1 polarization, the mRNA levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were markedly upregulated, the expression of CD86 was enhanced, and the secretions of IL-1β and TNF-α in THP-1 Mφ were higher than those in PBMC Mφ. After M2 polarization, the mRNA levels of mannose receptor C-type 1 and interleukin-10 increased, and the expression of CD206 was upregulated in PBMC Mφ. However, no difference was observed in THP Mφ. Conclusion THP-1 is appropriate for M1 polarization, and PBMC is appropriate for M2 polarization.

Key words: macrophage polarization, human monocyte cell, peripheral blood mononuclear cell

CLC Number: 

  • Q25

TrendMD: 

Fig 1

Liquid was divided into three layers in density gradient centri-fugation"

Tab 1

Primer sequences of qRT-PCR analysis"

基因名称 引物序列（5’—3’）
IL-1β 正向：TGGCTTATTACAGTGGCAATGAGGATG
反向：TGTAGTGGTGGTCGGAGATTCGTAG
TNF-α 正向：AGCTGGTGGTGCCATCAGAGG
反向：TGGTAGGAGACGGCGATGCG
MRC1 正向：TTGGACGGATGGACGAGGAGTC
反向：CGGATCGTGTCTGGCATATGTAGC
IL-10 正向：GCCAAGCCTTGTCTGAGATGATCC
反向：GCTCCACGGCCTTGCTCTTG
GAPDH 正向：TGCACCACCAACTGCTTAGC
反向：GGCATGGACTGTGGTCATGAG

Fig 2

Morphological changes of Mφ from two cells sources inverted microscope"

Fig 3

Relative expression level of cytokine mRNA after Mφ polarization from two resources of cells"

Fig 4

The expression of polarization markers CD86 and CD206 from Mφ"

Tab 2

The expression levels of cytokines in Mφ supernatant from two resources of cells pg·mL-1"

细胞因子 对照组（静息状态） LPS+IFN-γ刺激 IL-4刺激
THP-1 PBMC THP-1 PBMC THP-1 PBMC
IL-1β 58.43±1.83 56.25±1.74 591.35±5.67 551.73±8.51 62.58±1.97 60.72±1.81
TNF-α 41.57±2.35 39.16±2.27 436.91±10.72 338±13.51 45.37±2.48 41.42±2.71
IL-6 28.73±1.98 47.39±2.31 307.59±9.53 429.86±11.62 36.26±1.76 49.98±2.74
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