Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model

Introduction: Autologous mononuclear cells (MNCs) have been used in vascular regenerative therapy since the identification of endothelial progenitor cells (EPCs). However, the efficacy of autologous EPC therapy for diseases such as diabetes and connective tissue disorders is limited due to deficienc...

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Main Authors: Satomi Furukawa, Rie Hirano, Ai Sugawara, Satoshi Fujimura, Rica Tanaka
Format: Article
Language:English
Published: Elsevier 2024-06-01
Series:Regenerative Therapy
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352320424001159
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author Satomi Furukawa
Rie Hirano
Ai Sugawara
Satoshi Fujimura
Rica Tanaka
author_facet Satomi Furukawa
Rie Hirano
Ai Sugawara
Satoshi Fujimura
Rica Tanaka
author_sort Satomi Furukawa
collection DOAJ
description Introduction: Autologous mononuclear cells (MNCs) have been used in vascular regenerative therapy since the identification of endothelial progenitor cells (EPCs). However, the efficacy of autologous EPC therapy for diseases such as diabetes and connective tissue disorders is limited due to deficiencies in the number and function of EPCs. To address this, we developed a novel RE-01 cells that enriches pro-angiogenic cells from peripheral blood MNCs (PBMNCs). Methods: PBMNCs were collected from healthy volunteers following ethical guidelines. RE-01 cells were cultured in the presence of specific growth factors for 5 days without media change. Flow cytometry was used to analyze cell surface markers. Tube formation assays, EPC culture assays, and mRNA analysis were performed to evaluate angiogenic potential. The efficacy of RE-01 cells upon transplantation into ischemic hind limbs of mice was evaluated. Results: RE-01 cells exhibited a significant increase in pro-angiogenic cells such as M2 macrophages and angiogenic T cells, in contrast to PBMNCs, while the number of inflammatory cells reduced. In vitro assays demonstrated the enhanced angiogenic abilities of RE-01 cells, supported by increased mRNA expression of angiogenesis-related cytokines. In vivo studies using mouse ischemic hind limb models have shown that blood flow and angiogenesis improved following RE-01 cell transplantation. Transplantations for 3 consecutive days significantly improved the number of pericyte-recruited vessels in the severely ischemic hind limbs of mice. Conclusions: RE-01 cells showed promising results in enhancing angiogenesis and arteriogenesis, possibly owing to the presence of M2 macrophages and angiogenic T cells. These cells also demonstrated anti-fibrotic effects. The efficacy of RE-01 cells has been confirmed in mouse models, suggesting their potential for treating ischemic vascular diseases. Clinical trials are planned to validate the safety and efficacy of RE-01 cell therapy in patients with connective tissue disease and unhealed ulcers. We hope that this new RE-01 cell therapy will prevent many patients from undergoing amputation.
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spelling doaj-art-aa8634e8d477442892d043a8aeb08a392025-08-20T02:48:58ZengElsevierRegenerative Therapy2352-32042024-06-012629930710.1016/j.reth.2024.06.009Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb modelSatomi Furukawa0Rie Hirano1Ai Sugawara2Satoshi Fujimura3Rica Tanaka4Division of Regenerative Therapy, Juntendo University Graduates School of Medicine, Tokyo, Japan; Center for Genomic and Regenerative Medicine, Juntendo University Graduate School of Medicine, Tokyo, JapanDivision of Regenerative Therapy, Juntendo University Graduates School of Medicine, Tokyo, Japan; ReEir. Inc., Tokyo, JapanDivision of Regenerative Therapy, Juntendo University Graduates School of Medicine, Tokyo, Japan; Intractable Disease Research Center, Juntendo University Graduates School of Medicine, Tokyo, JapanDivision of Regenerative Therapy, Juntendo University Graduates School of Medicine, Tokyo, Japan; Intractable Disease Research Center, Juntendo University Graduates School of Medicine, Tokyo, JapanDivision of Regenerative Therapy, Juntendo University Graduates School of Medicine, Tokyo, Japan; Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine, Tokyo, Japan; Center for Genomic and Regenerative Medicine, Juntendo University Graduate School of Medicine, Tokyo, Japan; Intractable Disease Research Center, Juntendo University Graduates School of Medicine, Tokyo, Japan; ReEir. Inc., Tokyo, Japan; Corresponding author. Juntendo University Graduate School of Medicine, Division of Regenerative Therapy, 2-1-1 Hongo, Bunkyo-ku, Tokyo, 113-8421, Japan.Introduction: Autologous mononuclear cells (MNCs) have been used in vascular regenerative therapy since the identification of endothelial progenitor cells (EPCs). However, the efficacy of autologous EPC therapy for diseases such as diabetes and connective tissue disorders is limited due to deficiencies in the number and function of EPCs. To address this, we developed a novel RE-01 cells that enriches pro-angiogenic cells from peripheral blood MNCs (PBMNCs). Methods: PBMNCs were collected from healthy volunteers following ethical guidelines. RE-01 cells were cultured in the presence of specific growth factors for 5 days without media change. Flow cytometry was used to analyze cell surface markers. Tube formation assays, EPC culture assays, and mRNA analysis were performed to evaluate angiogenic potential. The efficacy of RE-01 cells upon transplantation into ischemic hind limbs of mice was evaluated. Results: RE-01 cells exhibited a significant increase in pro-angiogenic cells such as M2 macrophages and angiogenic T cells, in contrast to PBMNCs, while the number of inflammatory cells reduced. In vitro assays demonstrated the enhanced angiogenic abilities of RE-01 cells, supported by increased mRNA expression of angiogenesis-related cytokines. In vivo studies using mouse ischemic hind limb models have shown that blood flow and angiogenesis improved following RE-01 cell transplantation. Transplantations for 3 consecutive days significantly improved the number of pericyte-recruited vessels in the severely ischemic hind limbs of mice. Conclusions: RE-01 cells showed promising results in enhancing angiogenesis and arteriogenesis, possibly owing to the presence of M2 macrophages and angiogenic T cells. These cells also demonstrated anti-fibrotic effects. The efficacy of RE-01 cells has been confirmed in mouse models, suggesting their potential for treating ischemic vascular diseases. Clinical trials are planned to validate the safety and efficacy of RE-01 cell therapy in patients with connective tissue disease and unhealed ulcers. We hope that this new RE-01 cell therapy will prevent many patients from undergoing amputation.http://www.sciencedirect.com/science/article/pii/S2352320424001159Peripheral blood mononuclear cellCell therapyVasculogenesis
spellingShingle Satomi Furukawa
Rie Hirano
Ai Sugawara
Satoshi Fujimura
Rica Tanaka
Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model
Regenerative Therapy
Peripheral blood mononuclear cell
Cell therapy
Vasculogenesis
title Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model
title_full Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model
title_fullStr Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model
title_full_unstemmed Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model
title_short Novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model
title_sort novel cell therapy with ex vivo cultured peripheral blood mononuclear cells significantly impacts angiogenesis in the murine ischemic limb model
topic Peripheral blood mononuclear cell
Cell therapy
Vasculogenesis
url http://www.sciencedirect.com/science/article/pii/S2352320424001159
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