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Early injection of human adipose tissue-derived mesenchymal stem cell after inflammation ameliorates dextran sulfate sodium-induced colitis in mice through the induction of M2 macrophages and regulatory T cells

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Abstract

Inflammatory bowel diseases (IBDs) are sometimes refractory to current therapy or associated with severe adverse events during immunosuppressive therapy; thus, new therapies are urgently needed. Recently, mesenchymal stem cells (MSCs) have attracted attention based on their multitude of functions including anti-inflammatory effects. However, proper timing of MSC therapy and the mechanisms underlying the therapeutic effects of MSCs on colitis are not fully elucidated. Human adipose tissue-derived mesenchymal stem cells (hAdMSCs; 1 × 106) were administrated via the tail vein on day 3 (early) or 11 (delayed) using a 7-day dextran sulfate sodium (DSS)-induced mouse model of colitis. The effects were evaluated based on colon length, disease activity index (DAI) and histological score. Cytokine-encoding mRNA levels T cells and macrophages were evaluated by real-time PCR and flow cytometry. Regarding the timing of administration, early (day 3) injection significantly ameliorated DSS-induced colitis in terms of both DAI and histological score, compared to those parameters with delayed (day 11) injection. With early cell injection, the tissue mRNA levels of anti-inflammatory cytokine genes (Il10, Tgfb) increased, whereas those of inflammatory cytokine genes (Il6, Tnfa and Il17a) decreased significantly. Regarding the associated mechanism, hAdMSCs suppressed T cell proliferation and activation in vitro, increased the number of regulatory T cells in vivo and changed the polarity of macrophages (into the anti-inflammatory M2 phenotype) in vitro. Timing of injection is critical for the effective therapeutic effects of hAdMSCs. Furthermore, part of the associated mechanism includes T cell activation and expansion and altered macrophage polarization.

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Acknowledgments

The authors thank Takao Tsuchida (Department of Gastroenterology and Hepatology, Graduate School of Medical and Dental Science, Niigata University) for his cooperation.

Financial support

This work was supported by InterStem Co., Ltd.

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Authors and Affiliations

  1. Division of Gastroenterology and Hepatology, Graduate school of medical and dental sciences, Niigata University, 1-757 Asahimachi-dori, Chuo-ku, Niigata, 951-8510, Japan

    Yuzo Kawata, Atsunori Tsuchiya, Satoshi Seino, Yusuke Watanabe, Yuichi Kojima, Shunzo Ikarashi, Kentaro Tominaga, Junji Yokoyama, Satoshi Yamagiwa & Shuji Terai

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  1. Yuzo Kawata
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  2. Atsunori Tsuchiya
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  3. Satoshi Seino
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  4. Yusuke Watanabe
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  5. Yuichi Kojima
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  6. Shunzo Ikarashi
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Correspondence to Atsunori Tsuchiya or Shuji Terai.

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ST received research funding from InterStem Co., Ltd.

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Supplemental Figure 1

Evaluation of the therapeutic effect of human adipose tissue-derived mesenchymal stem cells (hAdMSCs) in a mouse model of dextran sulfate sodium-induced colitis, after intra-peritoneal administration on day 3 after initiation. (a and b) Prevention of colon length shortening and improved disease activity index (DAI) scores were observed in the group intra-peritoneally administered hAdMSCs on day 3 and sacrificed on day 21 (N = 18 mice; control, 8 mice, injected, 10 mice). (c) Histological analysis. Significant improvement in histological sores was not observed in this group. (d) Analysis of mRNA expression changes in the middle colon in this group, compared to levels in the untreated group (*P < 0.05). N.S., not significant. (PNG 183 kb)

High Resolution Image (TIF 299 kb)

Supplemental Figure 2

Evaluation of the therapeutic effect of human adipose tissue-derived mesenchymal stem cells (hAdMSCs) in a mouse model of dextran sulfate sodium-induced colitis, after administration using the tail vein on day 7 after initiation. (a and b) Prevention of colon length shortening was observed; however, improved disease activity index (DAI) scores were not observed in the group administered hAdMSCs using the tail vein on day 7 and sacrificed on day 21 (N = 13 mice; control, 6 mice, injected, 7 mice). (c) Histological analysis. Significant improvement in histological scores was not observed in this group. (d) Analysis of mRNA expression changes in the middle colon in this group, compared to levels in the untreated group (*P < 0.05). N.S., not significant. (PNG 176 kb)

High Resolution Image (TIF 281 kb)

Supplemental Figure 3

Effect of human adipose tissue-derived mesenchymal stem cells (hAdMSCs) on macrophages. Evaluation of CD206 (M2) macrophages by flow cytometry (a-c). The frequency of CD206-positive cells/F4/80-positive cells significantly increased in the hAdMSC-injected group compared to that in the control group (*P < 0.05). (PNG 322 kb)

High Resolution Image (TIF 446 kb)

Supplemental Table 1

List of primers used for real-time PCR (DOC 94 kb)

Supplemental Table 2

List of antibodies used for immunostaining (DOC 77 kb)

Supplemental Table 3

List of antibodies used for flow cytometry (DOCX 12 kb)

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Kawata, Y., Tsuchiya, A., Seino, S. et al. Early injection of human adipose tissue-derived mesenchymal stem cell after inflammation ameliorates dextran sulfate sodium-induced colitis in mice through the induction of M2 macrophages and regulatory T cells. Cell Tissue Res 376, 257–271 (2019). https://doi.org/10.1007/s00441-018-02981-w

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  • DOI: https://doi.org/10.1007/s00441-018-02981-w

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