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

Autor: Manabu Takewaki, Kazuhisa Ouhara, Tomoyuki Iwata, Nao Komatsu, Hidemi Kurihara, Tsuyoshi Fujita, Mikihito Kajiya, Souta Motoike, Kei Takeshita, Katsuhiro Takeda, Noriyoshi Mizuno
Rok vydání: 2016
Předmět:
0301 basic medicine
Male
Bone Regeneration
medicine.medical_treatment
T-Lymphocytes
Medicine (miscellaneous)
Ascorbic Acid
Mice
SCID
IDO
Cell therapy
Extracellular matrix
Mice
Mice
Inbred NOD
lcsh:QD415-436
C-MSC
IFN-γ
Cells
Cultured
lcsh:R5-920
Cell biology
medicine.anatomical_structure
Molecular Medicine
Xenotransplantation
Stem cell
lcsh:Medicine (General)
T cell
Transplantation
Heterologous
Bone Marrow Cells
Biology
Mesenchymal Stem Cell Transplantation
Biochemistry
Genetics and Molecular Biology (miscellaneous)
lcsh:Biochemistry
03 medical and health sciences
Interferon-gamma
Downregulation and upregulation
medicine
Animals
Humans
Indoleamine-Pyrrole 2
3
-Dioxygenase
Bone regeneration
Cell Proliferation
Research
Mesenchymal stem cell
Skull
Mesenchymal Stem Cells
Cell Biology
HLA-DR Antigens
Mice
Inbred C57BL
Disease Models
Animal
030104 developmental biology
Immunology
Leukocytes
Mononuclear
Zdroj: Stem Cell Research & Therapy
Stem Cell Research & Therapy, Vol 8, Iss 1, Pp 1-14 (2017)
ISSN: 1757-6512
Popis: Background 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. Methods 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. Results 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. Conclusions 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. Electronic supplementary material The online version of this article (doi:10.1186/s13287-017-0550-1) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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