Effect of human umbilical cord blood mesenchymal stem cells on the transformation of M2 macrophages from mouse bone marrow

doi:10.3969/j.issn.2095-4344.1778

Abstract

Abstract:

BACKGROUND: Mesenchymal stem cells have been used in the treatment of a variety of diseases. They may produce biological effects by promoting the conversion of macrophages to M2 subtypes, but it is unclear whether these effects are dose- and time-sensitive, as well as whether intercellular direct contact is involved in the cell transformation.
OBJECTIVE: To verify the effect of umbilical cord blood mesenchymal stem cells (hUC-MSCs) on the transformation of M2 subtypes of macrophages, and to investigate the dose- and time-dependent effects of hUC-MSCs on the transformation of M2 subtypes of macrophages.
METHODS: Monocytes were extracted from mouse bone marrow and induced by macrophage colony-stimulating factor to produce M0 macrophages (F4/80⁺, CD11B⁺). M0 macrophages were implanted into Transwen plates or 6-well culture plates at a density of 1×10⁶/well, and co-cultured directly or indirectly with hUC-MSCs at different concentrations (0, 2×10⁵, 4×10⁵) in complete medium containing lipopolysaccharide (100 μg/L) and interferon-γ (20 μg/L) for 24 and 48 hours. The morphological changes of macrophages were observed under direct microscope. The number of M2 macrophages (CD206⁺, CD11C^-) was detected by flow cytometry. The levels of interleukin-10, interleukin-6 and interleukin-1β in the cell culture supernatant were measured by ELISA.
RESULTS AND CONCLUSION: (1) After co-cultured with hUC-MSCs, the macrophages were elongated in cell antennae and the funicular cells, like M2 macrophages, appeared. (2) After 24 and 48 hours of direct or indirect co-culture with hUC-MSCs, the percentage of M2 macrophages was significantly increased (P < 0.05). (3) The percentage of M2 macrophages insignificantly differed after direct or indirect co-culture with low- and high-concentration hUC-MSCs. (4) The percentage of M2 macrophages was significantly higher after 48 hours co-culture than after 24 hours co-culture (P < 0.05). (5) After direct or indirect co-culture with hUC-MSCs, the levels of interleukin 6 and interleukin-1β in the cell supernatant were decreased significantly, while the interleukin-10 level increased significantly (P < 0.05). (6) The interleukin-10 level was significantly increased after 48 hours co-culture relative to that after 24 hours co-culture. In conclusion, hUC-MSCs can promote the transformation of M2 macrophages in a time-dependent but not concentration-dependent manner.

Key words: human umbilical cord blood mesenchymal stem cells, macrophages, macrophage M1/M2 subtype, M2 subtype transformation, inflammatory response, direct co-culture, indirect co-culture, National Natural Science Foundation of China

CLC Number:

Chen Bingquan, Peng Yi, Xiao Yi, Peng Zhiyong, Zhao Jiling, Yu Guolong. Effect of human umbilical cord blood mesenchymal stem cells on the transformation of M2 macrophages from mouse bone marrow[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 3987-3992.

Figures/Tables 3

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