Xenotransplantation of interferon-gamma-pretreated clumps of a human mesenchymal stem cell/extracellular matrix complex induces mouse calvarial bone regeneration.
Xenotransplantation of interferon-gamma-pretreated clumps of a human mesenchymal stem cell/extracellular matrix complex induces mouse calvarial bone regeneration.
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DOI：
10.1186/s13287-017-0550-1
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发表时间：
2017-04-26
影响因子：
7.5
通讯作者：
Kurihara H
中科院分区：
文献类型：
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作者：
Takeshita K;Motoike S;Kajiya M;Komatsu N;Takewaki M;Ouhara K;Iwata T;Takeda K;Mizuno N;Fujita T;Kurihara H
间充质干细胞（MSC）/细胞外基质（ECM）复合物（C-MSC）的三维培养团块由细胞和自身产生的ECM组成。C-MSC可以在体外调节细胞功能，并以ECM为细胞支架诱导成功的骨再生。增强C-MSCs的免疫调节能力，可以改善同种异体特异性免疫应答，可能有助于开发有益的“现成”细胞疗法用于组织再生。据报道，干扰素（IFN）-γ通过上调免疫调节酶IDO来刺激MSC的免疫抑制特性。因此，本研究的目的是探讨IFN-γ对C-MSC免疫调节能力的影响，并在小鼠颅骨缺损模型中检测C-MSC或IFN-γ预处理的C-MSC（C-MSCγ）异种移植物的骨再生活性。将人骨髓来源的MSC以2.0 × 105个细胞/孔的密度接种到24孔板中，并在补充有50 μg/mL L-抗坏血酸的生长培养基中培养4天。为了获得C-MSC，使用微量移液器尖端刮擦在细胞片上形成的汇合细胞，然后撕下。将细胞片卷起以形成圆形细胞团。用IFN-γ和IDO刺激C-MSC表达，在体外评估免疫抑制能力和免疫表型。将C-MSC和C-MSCγ分别移植到免疫活性和免疫缺陷小鼠颅骨缺损模型中。IFN-γ刺激C-MSC中的IDO表达。C-MSCγ而非C-MSC减弱CD 3/CD 28诱导的T细胞增殖，并且其抑制作用被IDO抑制剂逆转。C-MSCγ可上调HLA-DR的表达，但未检测到其共刺激分子CD 86。将C-MSCγ异种移植到免疫活性小鼠颅骨缺损中诱导骨再生，而C-MSC异种移植失败并诱导移植区域中的T细胞浸润。另一方面，C-MSC和C-MSCγ异种移植到免疫缺陷小鼠中引起骨再生。C-MSCγ在体外通过上调IDO活性发挥免疫调节特性，异种移植可减弱异种反应性宿主免疫应答，从而诱导小鼠骨再生。因此，C-MSCγ可能成为一种有前途的新型同种异体骨细胞治疗骨再生。本文的在线版本（doi：10.1186/s13287-017-0550-1）包含补充材料，可供授权用户使用。
Three-dimensional cultured clumps of a mesenchymal stem cell (MSC)/extracellular matrix (ECM) complex (C-MSC) consists of cells and self-produced ECM. C-MSC can regulate the cellular function in vitro and induce successful bone regeneration using ECM as a cell scaffold. Potentiating the immunomodulatory capacity of C-MSCs, which can ameliorate the allo-specific immune response, may be helpful in developing beneficial “off-the-shelf” cell therapy for tissue regeneration. It is well reported that interferon (IFN)-γ stimulates the immunosuppressive properties of MSC via upregulation of the immunomodulatory enzyme IDO. Therefore, the aim of this study was to investigate the effect of IFN-γ on the immunomodulatory capacity of C-MSC in vitro and to test the bone regenerative activity of C-MSC or IFN-γ-pretreated C-MSC (C-MSCγ) xenografts in a mice calvarial defect model. Human bone marrow-derived MSCs were seeded at a density of 2.0 × 105 cells/well into 24-well plates and cultured with growth medium supplemented with 50 μg/mL L-ascorbic acid for 4 days. To obtain C-MSC, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and were then torn off. The cellular sheet was rolled to make a round clump of cells. C-MSC was stimulated with IFN-γ and IDO expression, immunosuppressive capacity, and immunophenotype were evaluated in vitro. Moreover, C-MSC or C-MSCγ was xenotransplanted into immunocompetent or immunodeficient mice calvarial defect models without artificial scaffold, respectively. IFN-γ stimulated IDO expression in C-MSC. C-MSCγ, but not C-MSC, attenuated CD3/CD28-induced T cell proliferation and its suppressive effect was reversed by an IDO inhibitor. C-MSCγ showed upregulation of HLA-DR expression, but its co-stimulatory molecule, CD86, was not detected. Xenotransplantation of C-MSCγ into immunocompetent mice calvarial defect induced bone regeneration, whereas C-MSC xenograft failed and induced T cell infiltration in the grafted area. On the other hand, both C-MSC and C-MSCγ xenotransplantation into immunodeficient mice caused bone regeneration. Xenotransplantation of C-MSCγ, which exerts immunomodulatory properties via the upregulation of IDO activity in vitro, may attenuate xenoreactive host immune response, and thereby induce bone regeneration in mice. Accordingly, C-MSCγ may constitute a promising novel allograft cell therapy for bone regeneration. The online version of this article (doi:10.1186/s13287-017-0550-1) contains supplementary material, which is available to authorized users.