Clumps of a mesenchymal stromal cell/extracellular matrix complex can be a novel tissue engineering therapy for bone regeneration

Kittaka, Mizuho; Kajiya, Mikihito; Shiba, Hideki; Manabu, Takewaki; Takeshita, Kei; Khung, Rathvisal; Fujita, Takako; Iwata, Takeo; Nguyen, Truong Quoc; Ouhara, Kazuhisa; Takeda, Katsuhiro; Fujita, Tsuyoshi; Kurihara, Hidemi

doi:10.1016/j.jcyt.2015.01.007

Cited by 44 publications

(75 citation statements)

References 29 publications

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“…We have previously demonstrated that syngenic rat C-MSC transplantation into a rat calvarial defect model caused successful bone regeneration [12]. In this present study, human C-MSC xenograft into immunocompetent mice calvarial defect induced T cell infiltration and failed to induce bone regeneration, whereas xenotransplantation of C-MSCγ, which showed higher IDO expression level and T cell suppressive capacity in vitro, caused bone healing.…”

Section: Discussionmentioning

confidence: 44%

“…Indeed, recent studies also reported the effectiveness of the scaffold-free fabrication of stem cell constructs for bone regenerative therapy [13, 14]. More importantly, we have discovered that the implantation of C-MSCs, cultured with osteoinductive medium in vitro, exerts more effective bone regenerative properties in a rat calvarial defect model [12]. This finding implied that C-MSCs cellular function can be regulated in vitro before transplantation.…”

Section: Introductionmentioning

confidence: 66%

“…The cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Sigma-Aldrich, Steinheim, Germany) supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA), 100 U/mL penicillin (Sigma-Aldrich), and 100 μg/mL streptomycin (Sigma-Aldrich), and then C-MSCs were prepared as previously reported with minor modifications [12]. Briefly, MSCs were seeded at a density of 2.0 × 10 5 cells/well into 24-well plates (Corning, Corning, NY, USA) and cultured with high-glucose DMEM (Sigma-Aldrich) supplemented with 10% fetal FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 50 μg/ml L-ascorbic acid (Sigma-Aldrich) for 4 days.…”

Section: Methodsmentioning

confidence: 99%

“…Very recently, we generated clumps of an MSC/extracellular matrix (ECM) complex (C-MSC), which consisted of cells and self-produced ECM [12]. C-MSC had good handleability and can be transplanted into bony lesions without artificial scaffold.…”

Section: Introductionmentioning

confidence: 99%

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Xenotransplantation of interferon-gamma-pretreated clumps of a human mesenchymal stem cell/extracellular matrix complex induces mouse calvarial bone regeneration

et al. 2017

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BackgroundThree-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.MethodsHuman 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.ResultsIFN-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.ConclusionsXenotransplantation 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0550-1) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 44%

Section: Introductionmentioning

confidence: 66%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Xenotransplantation of interferon-gamma-pretreated clumps of a human mesenchymal stem cell/extracellular matrix complex induces mouse calvarial bone regeneration

et al. 2017

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“…A variety of gelable materials and relevant cell types have been used in this system, and they are being investigated for advanced biomaterial-based cell delivery. More broadly, modular approaches to creating engineered tissue are emerging as a promising approach to creating complex structures [71–76]. Such methods have advantages over conventional scaffold-based techniques [64, 72], and new methods are being developed at a rapid rate.…”

Section: Discussionmentioning

confidence: 99%

Collagen Type II enhances chondrogenic differentiation in agarose-based modular microtissues

et al. 2016

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Background Cell-based therapies have made an impact on the treatment of osteoarthritis, however the repair and regeneration of thick cartilage defects is an important and growing clinical problem. Next-generation therapies that combine cells with biomaterials may provide improved outcomes. We have developed modular microenvironments that mimic the composition of articular cartilage as a delivery system for consistently differentiated cells. Methods Human bone marrow-derived mesenchymal stem cells (MSC) were embedded in modular microbeads consisting of agarose (AG) supplemented with 0%, 10%, and 20% collagen Type II (COL-II) using a water-in-oil emulsion technique. AG and AG/COL-II microbeads were characterized in terms of their structural integrity, size distribution, and protein content. The viability of embedded MSC and their ability to differentiate into osteogenic, adipogenic and chondrogenic lineages over three weeks in culture were also assessed. Results Microbeads made with <20% COL-II were robust, generally spheroidal in shape and 80 ± 10 µm in diameter. MSC viability in microbeads was consistently high over a week in culture, while viability in corresponding bulk hydrogels decreased with increasing COL-II content. Osteogenic differentiation of MSC was modestly supported in both AG and AG/COL-II microbeads, while adipogenic differentiation was strongly inhibited in COL-II containing microbeads. Chondrogenic differentiation of MSC was clearly promoted in microbeads containing COL-II, compared to pure AG matrices. Conclusions Inclusion of collagen Type II in agarose matrices in microbead format can potentiate chondrogenic differentiation of human MSC. Such compositionally tailored microtissues may find utility for cell delivery in next-generation cartilage repair therapies.

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Important Considerations in the Therapeutic Application of Stem Cells in Bone Healing and Regeneration

Elkhenany

Bourdo

Biris

et al. 2016

Stem Cells in Toxicology and Medicine

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Clumps of a mesenchymal stromal cell/extracellular matrix complex can be a novel tissue engineering therapy for bone regeneration

Cited by 44 publications

References 29 publications

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

Collagen Type II enhances chondrogenic differentiation in agarose-based modular microtissues

Important Considerations in the Therapeutic Application of Stem Cells in Bone Healing and Regeneration

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