Isolation of tumor endothelial cells from murine cancer

Taguchi, Kazuhiro; Onoe, Takashi; Yoshida, Tomoaki; Yamashita, Yoshihisa; Taniyama, Kiyomi; Ohdan, Hideki

doi:10.1016/j.jim.2018.11.005

Cited by 5 publications

(2 citation statements)

References 27 publications

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“…We had previously described a method for the isolation of highly pure, fresh TECs from tumors (24). Briefly, tumors were dissected from tumor-bearing mice and digested to single cells using Collagenase Type II (Worthington).…”

Section: Isolation Of Tecsmentioning

confidence: 99%

Tumor Endothelial Cell–Mediated Antigen-Specific T-cell Suppression via the PD-1/PD-L1 Pathway

Taguchi

Onoe

Yoshida

et al. 2020

Molecular Cancer Research

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Tumor endothelial cells (TEC) play multiple roles in the regional specialization of vascular structure and physiology. Because TECs in the tumor microenvironment come in contact with circulating immune cells, they might influence not only trafficking but also the antitumor cellular immune response. In a mouse tumor implantation model with B16 melanoma cells, TECs expressed MHC class II, costimulating molecules, and programmed death-ligand 1 (PD-L1), suggesting that they are antigen (Ag)-presenting cells with suppressive activity. Furthermore, TECs were able to take up and present tumor-derived ovalbumin (OVA) peptide on MHC class I molecules. In functional assays, B16-OVA tumor-derived TECs significantly suppressed the proliferation and Ag-specific cytotoxicity of OVA-specific CD8 þ T cells relative to those of B16 tumor-derived TECs. This suppressive activity required cell-cell contact and was abrogated by PD-L1 blockade. TECs impaired proinflammatory cytokine production of CD8 þ T cells, including IL2, TNFa, and IFNg. B16-OVA tumorderived TECs induced immunosuppressive CD4 þ T cells that suppressed OVA-specific CD8 þ T-cell proliferation via inhibitory cytokines, including IL10 and TGFb. Deficiency of PD-L1 in TECs, but not in hematopoietic cells, impaired suppression and apoptosis of tumor-infiltrating CD8 þ T cells, resulting in inhibition of tumor development in vivo model. These data suggest that TECs might regulate the immune response of tumor Ag-specific CD8 þ T cells via the PD-1/PD-L1 pathway and induce immune suppressive CD4 þ T cells in an Ag-specific manner, contributing to tumor immune evasion. Implications:The findings of this study might encourage the further development of novel anticancer therapies and strategies.

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Section: Isolation Of Tecsmentioning

confidence: 99%

Tumor Endothelial Cell–Mediated Antigen-Specific T-cell Suppression via the PD-1/PD-L1 Pathway

Taguchi

Onoe

Yoshida

et al. 2020

Molecular Cancer Research

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“…ECs grown in 2D and 3D cultures using matrigel, collagen, fibrinogen-thrombin gels, chicken membranes, agarose transplants harboring ECs, delivery of ex vivo cultured myoblasts into rodent ears are widely used models to study angiogenesis and associated cellular and molecular mechanisms (Philippova et al, 2006;Bray et al, 2017;Norrby et al, 2006). Human umbilical vein endothelial cells (HUVEC) exposed to hypoxia or conditioned medium of tumor cells (Burrows et al,1995;Hellebrekers et al, 2007;Motz et al, 2014) and EC isolated from nude mice xenograft models (Matsuda et al, 2010;Taguchi et al, 2019;Xiao et al,2015) are also frequently used as tumor angiogenesis models. However, these models possess limitations as pathologically, tumor EC show significant heterogeneity and are highly dependent on the microenvironment of the host (Hida et al, 2016;Palma et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Human breast tumor derived endothelial cells exhibit distinct biological properties

Hegde

Bhat

Guruprasad

et al. 2021

Biology of the Cell

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Background Information: Excessive angiogenesis characterized by leaky, tortuous, and chaotic vasculature is one of the hallmarks of cancers and is significantly correlated to poor prognosis. Disorganized angiogenesis leads to poor perfusion of anti-cancer drugs and limits access to immune cells. Hence, impeding angiogenesis is one of the attractive therapeutic targets to inhibit progression and metastasis in several solid tumors including breast. Results:We have developed a robust and reproducible method for isolating and ex vivo culture of endothelial cells (EC) derived from non-malignant (Endo-N) and malignant (Endo-T) part from clinically characterized human breast tumors. RT-PCR and immunoblotting analysis indicated that these cells exhibited expression of endothelial specific genes such as PECAM-1 (CD31), Endoglin (CD105), eNOS, VE-cadherin, VCAM1, and MCAM. Vasculogenic mimicry and contamination of progenitor EC recruited in tumors was ruled out by absence of CD133 expression and normal karyotype. Both the cell types showed stable expression of CD31 and CD105 up to seven passages. Furthermore, compared to Endo-N cells, Endo-T cells showed (a) constitutively increased proliferation marked by nearly 36% of cells in mitotic phase, (b) requirement of glutamine for cell survival, (c) pro-migratory phenotype, (d) produced increased number of sprouts in 3D cultures, and (e) resistance to sorafenib. Conclusion: Tumor derived EC showed distinct biological properties compared to normal breast EC. Significance: Our method for isolating endothelial cell types from human breast tumors may be explored to (a) understand cellular and molecular mechanisms, (b) screen anti-angiogenic molecules, and (c) formulate organoid cultures to develop personalized medicine facilitating better clinical management of breast cancers.

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Rebuilding the microenvironment of primary tumors in humans: a focus on stroma

Mun,

Lee,

Kim

2024

Exp Mol Med

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Conventional tumor models have critical shortcomings in that they lack the complexity of the human stroma. The heterogeneous stroma is a central compartment of the tumor microenvironment (TME) that must be addressed in cancer research and precision medicine. To fully model the human tumor stroma, the deconstruction and reconstruction of tumor tissues have been suggested as new approaches for in vitro tumor modeling. In this review, we summarize the heterogeneity of tumor-associated stromal cells and general deconstruction approaches used to isolate patient-specific stromal cells from tumor tissue; we also address the effect of the deconstruction procedure on the characteristics of primary cells. Finally, perspectives on the future of reconstructed tumor models are discussed, with an emphasis on the essential prerequisites for developing authentic humanized tumor models.

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Isolation of tumor endothelial cells from murine cancer

Cited by 5 publications

References 27 publications

Tumor Endothelial Cell–Mediated Antigen-Specific T-cell Suppression via the PD-1/PD-L1 Pathway

Tumor Endothelial Cell–Mediated Antigen-Specific T-cell Suppression via the PD-1/PD-L1 Pathway

Human breast tumor derived endothelial cells exhibit distinct biological properties

Rebuilding the microenvironment of primary tumors in humans: a focus on stroma

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