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https://www.arca.fiocruz.br/handle/icict/66748
CELLULAR IN VITRO RESPONSES INDUCED BY HUMAN MESENCHYMAL STEM/STROMAL CELL-DERIVED EXTRACELLULAR VESICLES OBTAINED FROM SUSPENSION CULTURE
Angiogênese
Polarização de macrófagos
Células-tronco
Reparação de ferida
Estromais mesenquimais derivadas de tecido adiposo
Adipose tissue-derived mesenchymal stem
Stromal cells
Wound repair
Angiogenesis
Macrophage polarization
Author
Affilliation
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Hospital Israelita Albert Einstein. São Paulo, SP, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Plataforma de Microscopia Eletrônica e Confocal. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Plataforma de Microscopia Eletrônica e Confocal. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Hospital Israelita Albert Einstein. São Paulo, SP, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Plataforma de Microscopia Eletrônica e Confocal. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Plataforma de Microscopia Eletrônica e Confocal. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-tronco. Curitiba, PR, Brasil.
Abstract
Mesenchymal stem/stromal cells (MSCs) and their extracellular vesicles (MSC-EVs) have been described to have important roles in tissue regeneration, including tissue repair, control of inflammation, enhancing angiogenesis, and regulating extracellular matrix remodeling. MSC-EVs have many advantages for use in regeneration therapies such as facility for dosage, histocompatibility, and low immunogenicity, thus possessing a lower possibility of rejection. In this work, we address the potential activity of MSC-EVs isolated from adipose-derived MSCs (ADMSC-EVs) cultured on cross-linked dextran microcarriers, applied to test the scalability and reproducibility of EV production. Isolated ADMSC-EVs were added into cultured human dermal fibroblasts (NHDF-1), keratinocytes (HaCat), endothelial cells (HUVEC), and THP-1 cell-derived macrophages to evaluate cellular responses (i.e., cell proliferation, cell migration, angiogenesis induction, and macrophage phenotype-switching). ADMSC viability and phenotype were assessed during cell culture and isolated ADMSC-EVs were monitored by nanotracking particle analysis, electron microscopy, and immunophenotyping. We observed an enhancement of HaCat proliferation; NHDF-1 and HaCat migration; endothelial tube formation on HUVEC; and the expression of inflammatory cytokines in THP-1-derived macrophages. The increased expression of TGF-β and IL-1β was observed in M1 macrophages treated with higher doses of ADMSC-EVs. Hence, EVs from microcarrier-cultivated ADMSCs are shown to modulate cell behavior, being able to induce skin tissue related cells to migrate and proliferate as well as stimulate angiogenesis and cause balance between pro- and antiinflammatory responses in macrophages. Based on these findings, we suggest that the isolation of EVs from ADMSC suspension cultures makes it possible to induce in vitro cellular responses of interest and obtain sufficient particle numbers for the development of in vivo concept tests for tissue regeneration studies.
Keywords in Portuguese
Vesículas extracelularesAngiogênese
Polarização de macrófagos
Células-tronco
Reparação de ferida
Estromais mesenquimais derivadas de tecido adiposo
Keywords
Extracellular vesiclesAdipose tissue-derived mesenchymal stem
Stromal cells
Wound repair
Angiogenesis
Macrophage polarization
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