Decellularization of placentas: establishing a protocol

Leonel, Luciano; Miranda, Carla M. F. C.; Coelho, Talya Moraes; Ferreira, Guilherme A S; Caãada, R.R.; Miglino, Maurizio; Lobo, Sonja Ellen

doi:10.1590/1414-431x20176382

Cited by 31 publications

(46 citation statements)

References 41 publications

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“…We developed an acellular biological scaffold derived from canine placenta preserving its architecture. The presence of important proteins in your constitution (collagen, laminin, and fibronectin) 31 contributed of being recellularized with YSVEGF cells. The genomic DNA analysis showed complete scaffold decellularization using SDS and Triton 11 ( Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Canine Placenta Recellularized Using Yolk Sac Cells with Vascular Endothelial Growth Factor

Fratini

Rigoglio

Matias

et al. 2018

BioResearch Open Access

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Regenerative medicine has been growing because of the emergent need for tissues/organs for transplants and restorative surgeries. Biological scaffolds are important tools to try to solve this problem. The one used in this reserach was developed by an acellular biological scaffold from canine placenta with a rich source of cellular matrix. After decellularization, the cellular matrix demonstrated structural preservation with the presence of important functional proteins such as collagen, fibronectin, and laminin. We used cells transduced with vascular endothelial growth factor (VEGF) to recellularize this scaffold. It was succeeded by seeding the cells in nonadherent plaques in the presence of the sterelized placenta scaffold. Cells were adhered to the scaffold when analyzed by immunocytochemistry and scanning electron microscopy, both showing sprouting of yolk sac VEGF (YSVEGF) cells. This recellularized scaffold is a promissory biomaterial for repairing injured areas where neovascularization is required.

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Section: Resultsmentioning

confidence: 99%

Canine Placenta Recellularized Using Yolk Sac Cells with Vascular Endothelial Growth Factor

Fratini

Rigoglio

Matias

et al. 2018

BioResearch Open Access

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“…Decellularized organ-derived extracellular matrix (dECM) has been used for many years in tissue engineering and regenerative medicine (Conklin et al, 2002;Dahl et al, 2003;Schenke-Layland et al, 2003). Decellularization of the extracellular matrix can be achieved through different physical, chemical or biological processes, including freeze/thaw cycles, use of organic detergents, and mild treatment with proteolytic enzymes (Crapo et al, 2011;Hrebikova et al, 2015;Keane et al, 2015;Leonel et al, 2017;Kawecki et al, 2018). Commonly, different processes are combined or used sequentially, so as to decrease the damage caused by each of them to the extracellular matrix but, at the same time, to potentiate the decellularization protocol (Crapo et al, 2011;Kawecki et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Molecular and Biomechanical Clues From Cardiac Tissue Decellularized Extracellular Matrix Drive Stromal Cell Plasticity

Liguori

Moraes

et al. 2020

Front. Bioeng. Biotechnol.

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Decellularized-organ-derived extracellular matrix (dECM) has been used for many years in tissue engineering and regenerative medicine. The manufacturing of hydrogels from dECM allows to make use of the pro-regenerative properties of the ECM and, simultaneously, to shape the material in any necessary way. The objective of the present project was to investigate differences between cardiovascular tissues (left ventricle, mitral valve, and aorta) with respect to generating dECM hydrogels and their interaction with cells in 2D and 3D. The left ventricle, mitral valve, and aorta of porcine hearts were decellularized using a series of detergent treatments (SDS, Triton-X 100 and deoxycholate). Mass spectrometry-based proteomics yielded the ECM proteins composition of the dECM. The dECM was digested with pepsin and resuspended in PBS (pH 7.4). Upon warming to 37 • C, the suspension turns into a gel. Hydrogel stiffness was determined for samples with a dECM concentration of 20 mg/mL. Adipose tissue-derived stromal cells (ASC) and a combination of ASC with human pulmonary microvascular endothelial cells (HPMVEC) were cultured, respectively, on and in hydrogels to analyze cellular plasticity in 2D and vascular network formation in 3D. Differentiation of ASC was induced with 10 ng/mL of TGF-β1 and SM22α used as differentiation marker. 3D vascular network formation was evaluated with confocal microscopy after immunofluorescent staining of PECAM-1. In dECM, the most abundant protein was collagen VI for the left ventricle and mitral valve and elastin for the aorta. The stiffness of the hydrogel derived from the aorta (6,998 ± 895 Pa) was significantly higher than those derived from the left ventricle (3,384 ± 698 Pa) and the mitral valve (3,233 ± 323 Pa) (One-way ANOVA, p = 0.0008). Aorta-derived dECM hydrogel drove non-induced (without TGF-β1) differentiation, while hydrogels derived from the left ventricle and mitral valve inhibited TGF-β1-induced differentiation. All hydrogels supported vascular network formation within 7 days of culture, but ventricular dECM hydrogel demonstrated more Liguori et al. Tissue-Specific Hydrogels Drive Cell Plasticity robust vascular networks, with thicker and longer vascular structures. All the three main cardiovascular tissues, myocardium, valves, and large arteries, could be used to fabricate hydrogels from dECM, and these showed an origin-dependent influence on ASC differentiation and vascular network formation.

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“…Neste trabalho, as MECs de músculo esquelético murino foram descelularizadas de acordo com o protocolo testado previamente em nosso laboratório, utilizando métodos físicos e químicos (congelação a -20°C e incubação em 1% SDS, 5 mM EDTA+ 50mM Tris e 1% Triton X-100). Enquanto que a MEC de placenta canina foi descelularizada conforme a metodologia descrita por Leonel et al (2017), utilizando métodos enzimáticos e químicos (1% SDS, 5 mM EDTA+ 0,05% Tripsina e 1% Triton X-100). Ambas as matrizes obtidas apresentaram aspecto translúcido e gelatinoso, sendo comprovada a ausência de células e a preservação da estrutura tridimensional.…”

Section: Discussionunclassified

<b>Avaliação de matrizes descelularizadas para regeneração muscular</b>

Miranda¹

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Agradeço a Deus Pai Todo-poderoso por sempre ter estado ao meu lado e pela oportunidade que Ele me deu de estar aqui, hoje, concluindo mais uma etapa de minha vida. Etapa árdua, mas que com muita perserverança eu consegui chegar até o final. Agradeço eternamente a Virgem Maria pela Sua proteção e intercessão perante a Deus. "Eu sei que é difícil esperar, mas Deus tem um tempo para agir e para curar. Só é preciso confiar!!". Aos meus tios Nonato, Pedrinho, Ismael, Manoel e Zeca e, especialmente; às minhas maravilhosas tias Marta, Teca, Marcelina, Mônica e Cristina; aos meus primos, David, Denzel, Mateus, Renata e Anderson; aos meus queridos avós Maria Leontina, Erothildes (in memoriam), Carvalho (in memoriam), e Raimundo Pedro (in memoriam): agradeço imensamente a família FIGUEIREDO pelo apoio, força, confiança e esperança no meu sucesso profissional. Essa caminhada não teria sido possível sem vocês, pois vocês são o meu alicerce! À minha mãe, Joana D´arc, à minha irmã, Socorro, e ao meu pai, Luis Carlos, pelo apoio, amor, carinho e dedicação durante toda a minha trajetória na universidade. Obrigada pelo apoio nos momentos difíceis e longe de casa. Sou eternamente grata! Agora sabemos que todo o esforço valeu, e sempre, valerá à pena! Ao Moysés Miranda, não poderia deixar de agradecer pela paciente compreensão com esta jornada de doutorado. Agradeço sinceramente por todo o convívio nestes 13 anos e por tudo o que vida nos propôs a ensinar durante essa trajetória. Gratidão eterna também aos seus pais Zuíla (in memoriam) e Brás (in memoriam), e irmãos, Rosa e Marcelo, por terem me recebido na família com muito carinho e estendido a mão quando precisei. À professora Maria Angélica Miglino, pela sua dedicação como professora e por acreditar e confiar no meu trabalho. Agradeço pelas oportunidades que me foram concedidas em aprimorar meus conhecimentos e à minha formação, os quais foram muito valiosos para a minha carreira profissional. À Dra Sonja Ellen Lobo, pela sua dedicação na orientação deste trabalho e por todo o ensinamento repassado durante estes 4 anos. Obrigada pela confiança em meu trabalho. Aos técnicos Rose Eli (Laboratório de Microscopia) e Ronaldo Agostinho (Laboratório de Histologia), agradeço por toda a ajuda no laboratório durante a execução deste trabalho. A Paula Fratini pela sua sempre solicitude e ao Rodrigo Barreto, agradeço a colaboração nas análises deste trabalho e, principalmente, pela sua sempre boa disponibilidade em ajudar e ensinar.

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Decellularization of placentas: establishing a protocol

Cited by 31 publications

References 41 publications

Canine Placenta Recellularized Using Yolk Sac Cells with Vascular Endothelial Growth Factor

Canine Placenta Recellularized Using Yolk Sac Cells with Vascular Endothelial Growth Factor

Molecular and Biomechanical Clues From Cardiac Tissue Decellularized Extracellular Matrix Drive Stromal Cell Plasticity

<b>Avaliação de matrizes descelularizadas para regeneração muscular</b>

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