記述言語：日本語   出版者・発行元：一般社団法人 日本移植学会  

<p>【目的】　肺は最低でも58種類もの多種多様な細胞種が存在し非常に複雑な相互作用で恒常性が維持され、ターンオーバーが非常に遅く、再生能力に乏しい臓器である。そのため、研究は他臓器に比べ後れ、呼吸器領域の再生医療はまだまだ開発段階である。呼吸器再生医療の実用化のためには、基礎研究の拡充とシーズの発掘が必要である。【方法】　当科で実施した呼吸器再生研究を振り返り問題点を整理し、これまでの研究成果から新しい方向性を見いだす。【結果】これまでに代替臓器の創出を目指し1）3Dバイオプリンティング技術による気管再生、2）生体臓器鋳型を抽出する脱細胞化技術による肺血管網作成やex vivo再生肺などの臓器再生研究を行い、細胞治療の開発を目指して3）脂肪組織由来間葉系幹細胞による拒絶抑制効果や肺胞血管バリア機能の修復効果の検証を小動物肺移植モデルにて行ってきた。肺移植の代替となる再生肺の作成には多くの課題がありまだまだ挑戦的であり、細胞治療では肺への細胞生着率の改善が問題である。ただし、幹細胞をはじめとする各種細胞が形成する細胞凝集塊（スフェロイド）はより機能的な細胞治療となりうる可能性があり、生体由来組織の臓器骨格をもつ高次構造の足場などは細胞治療のin vitro研究の場を提供しうる。【考察】　細胞凝集塊を用いた肺細胞治療の生着率を改善する研究を進めることで、再生医療の実用化へ繋げる。</p>

DOI： 10.11386/jst.59.supplement_s142_2

CiNii Research

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Organogenesis  

Decellularized scaffolds are promising biomaterials for tissue and organ reconstruction; however, strategies to effectively suppress the host immune responses toward these implants, particularly those without chemical crosslinking, remain warranted. Administration of mesenchymal stem cells is effective against immune-mediated inflammatory disorders. Herein, we investigated the effect of isogeneic abdominal adipose-derived mesenchymal stem/stromal cells (ADMSCs) on xenogeneic biomaterial-induced immunoreactions. Peripheral bronchi from pigs, decellularized using a detergent enzymatic method, were engrafted onto tracheal defects of Brown Norway (BN) rats. BN rats were implanted with native pig bronchi (Xenograft group), decellularized pig bronchi (Decellularized Xenograft), or Decellularized Xenograft and ADMSCs (Decellularized Xenograft+ADMSC group). In the latter group, ADMSCs were injected intravenously immediately post implantation. Harvested graft implants were assessed histologically and immunohistochemically. We found that acute rejections were milder in the Decellularized Xenograft and Decellularized Xenograft+ADMSC groups than in the Xenograft group. Mild inflammatory cell infiltration and reduced collagen deposition were observed in the Decellularized Xenograft+ADMSC group. Additionally, ADMSC administration decreased CD8+ lymphocyte counts but increased CD163+ cell counts. In the Decellularized Xenograft+ADMSC group, serum levels of vascular endothelial growth factor and IL-10 were elevated and tissue deposition of IgM and IgG was low. The significant immunosuppressive effects of ADMSCs illustrate their potential use as immunosuppressive agents for xenogeneic biomaterial-based implants.

DOI： 10.1080/15476278.2023.2212582

Open Access

Scopus

PubMed

記述言語：日本語   出版者・発行元：(一社)日本臓器保存生物医学会  

記述言語：日本語   出版者・発行元：(一社)日本臓器保存生物医学会  

記述言語：日本語   出版者・発行元：(一社)日本臓器保存生物医学会  

記述言語：日本語   出版者・発行元：(一社)日本臓器保存生物医学会  

記述言語：日本語   出版者・発行元：(株)北隆館  

がん医学や再生医学などの最先端生命医学において,これまで細胞内の物理化学的パラメーターが機能発現に及ぼす影響はほとんど解明されていない。例えば,がん医学における超初期がん細胞の発生・駆逐メカニズムや再生医学における生体内での幹細胞維持・分化メカニズムなどの喫緊の重要課題においても,計測,評価できているのはシグナル分子(タンパク質,RNAなど)についてのみであり,機能発現に重要な物理化学的パラメーター(温度,pH濃度,柔軟性など)は計測出来ないが故に評価されていない。しかし,近年,ナノ量子センサーを応用することで,これらの多機能同時計測・評価が可能になると期待されている。本論文では,筆者らが開発してきた最先端ナノ量子センサーや計測デバイスを詳細するとともに,その計測・評価結果について概説する。(著者抄録)

掲載種別：研究論文（学術雑誌）  

DOI： 10.3390/cells12101417

Open Access

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers Media SA  

Introduction: Sjögren syndrome (SS) is an autoimmune disease characterized by salivary gland (SG) destruction leading to loss of secretory function. A hallmark of the disease is the presence of focal lymphocyte infiltration in SGs, which is predominantly composed of T cells. Currently, there are no effective therapies for SS. Recently, we demonstrated that a newly developed therapy using effective-mononuclear cells (E-MNCs) improved the function of radiation-injured SGs due to anti-inflammatory and regenerative effects. In this study, we investigated whether E-MNCs could ameliorate disease development in non-obese diabetic (NOD) mice as a model for primary SS.

Methods: E-MNCs were obtained from peripheral blood mononuclear cells (PBMNCs) cultured for 7 days in serum-free medium supplemented with five specific recombinant proteins (5G culture). The anti-inflammatory characteristics of E-MNCs were then analyzed using a co-culture system with CD3/CD28-stimulated PBMNCs. To evaluate the therapeutic efficacy of E-MNCs against SS onset, E-MNCs were transplanted into SGs of NOD mice. Subsequently, saliva secretion, histological, and gene expression analyses of harvested SG were performed to investigate if E-MNCs therapy delays disease development.

Results: First, we characterized that both human and mouse E-MNCs exhibited induction of CD11b/CD206-positive cells (M2 macrophages) and that human E-MNCs could inhibit inflammatory gene expressions in CD3/CD28- stimulated PBMNCs. Further analyses revealed that Msr1-and galectin3-positive macrophages (immunomodulatory M2c phenotype) were specifically induced in E-MNCs of both NOD and MHC class I-matched mice. Transplanted E-MNCs induced M2 macrophages and reduced the expression of T cell-derived chemokine-related and inflammatory genes in SG tissue of NOD mice at SS-onset. Then, E-MNCs suppressed the infiltration of CD4-positive T cells and facilitated the maintenance of saliva secretion for up to 12 weeks after E-MNC administration.

Discussion: Thus, the immunomodulatory actions of E-MNCs could be part of a therapeutic strategy targeting the early stage of primary SS.

DOI： 10.3389/fbioe.2023.1144624

Open Access

PubMed

記述言語：日本語   出版者・発行元：(一社)日本移植学会  

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Bioengineering and Biotechnology  

Introduction: Two-dimensional cell cultures have contributed substantially to lung cancer research, but 3D cultures are gaining attention as a new, more efficient, and effective research model. A model reproducing the 3D characteristics and tumor microenvironment of the lungs in vivo, including the co-existence of healthy alveolar cells with lung cancer cells, is ideal. Here, we describe the creation of a successful ex vivo lung cancer model based on bioengineered lungs formed by decellularization and recellularization. Methods: Human cancer cells were directly implanted into a bioengineered rat lung, which was created with a decellularized rat lung scaffold reseeded with epithelial cells, endothelial cells and adipose-derived stem cells. Four human lung cancer cell lines (A549, PC-9, H1299, and PC-6) were applied to demonstrate forming cancer nodules on recellularized lungs and histopathological assessment were made among these models. MUC-1 expression analysis, RNA-seq analysis and drug response test were performed to demonstrate the superiority of this cancer model. Results: The morphology and MUC-1 expression of the model were like those of lung cancer in vivo. RNA sequencing revealed an elevated expression of genes related to epithelial-mesenchymal transition, hypoxia, and TNF-α signaling via NF-κB; but suppression of cell cycle-related genes including E2F. Drug response assays showed that gefitinib suppressed PC-9 cell proliferation equally well in the 3D lung cancer model as in 2D culture dishes, albeit over a smaller volume of cells, suggesting that fluctuations in gefitinib resistance genes such as JUN may affect drug sensitivity. Conclusions: A novel ex vivo lung cancer model was closely reproduced the 3D structure and microenvironment of the actual lungs, highlighting its possible use as a platform for lung cancer research and pathophysiological studies.

DOI： 10.3389/fbioe.2023.1179830

Open Access

Scopus

PubMed

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Cell Transplantation  

Cell therapy using mesenchymal stromal cells (MSCs) is being studied for its immunosuppressive effects. In organ transplantation, the amount of MSCs that accumulate in transplanted organs and other organs may differ depending on administration timing, which may impact their immunosuppressive effects. In vitro, adipose-derived mesenchymal stem cells (ADMSCs) suppress lymphocyte activation under cell-to-cell contact conditions. However, in vivo, it is controversial whether ADMSCs are more effective in accumulating in transplanted organs or in secondary lymphoid organs. Herein, we aimed to investigate whether the timing of ADMSC administration affects its immunosuppression ability in a rat lung transplantation model. In the transplantation study, rats were intramuscularly administered half the usual dose of tacrolimus (0.5 mg/kg) every 24 h after lung transplantation. ADMSCs (1 × 106) were administered via the jugular vein before (PreTx) or after (PostTx) transplantation. Cell tracking using quantum dots was performed. ADMSCs accumulated predominantly in the lung and liver; fewer ADMSCs were distributed in the grafted lung in the PreTx group than in the PostTx group. The rejection rate was remarkably low in the ADMSC-administered groups, particularly in the PostTx group. Serum tumor necrosis factor-α (TNF-α), interferon-γ, and interleukin (IL)-6 levels showed a greater tendency to decrease in the PreTx group than in the PostTx group. The proportion of regulatory T cells in the grafted lung 10 days after transplantation was higher in the PostTx group than in the PreTx group. PostTx administration suppresses rejection better than PreTx administration, possibly due to regulatory T cell induction by ADMSCs accumulated in the transplanted lungs, suggesting a mechanism different from that in heart or kidney transplantation that PreTx administration is more effective than PostTx administration. These results could help establish cell therapy using MSCs in lung transplantation.

DOI： 10.1177/09636897231207177

Open Access

Scopus

PubMed

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