3D Bioprinting of Murine Cortical Astrocytes for Engineering Neural-Like Tissue

Autor(es)

de Melo, Bruna A. G. ^[1] ; Cruz, Elisa M. ^[1] ; Ribeiro, Tais N. ^[1] ; Mundim, V, Mayara ; Porcionatto, Marimelia A. ^[2]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Biochem, Sao Paulo - Brazil

^[2] Mundim, Mayara, V, Univ Fed Sao Paulo, Escola Paulista Med, Dept Biochem, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS; n. 173 JUL 2021.
• Citações Web of Science: 0

Resumo

Astrocytes are glial cells with an essential role in the central nervous system (CNS), including neuronal support and functionality. These cells also respond to neural injuries and act to protect the tissue from degenerative events. In vitro studies of astrocytes' functionality are important to elucidate the mechanisms involved in such events and contribute to developing therapies to treat neurological disorders. This protocol describes a method to biofabricate a neural-like tissue structure rich in astrocytes by 3D bioprinting astrocytes-laden bioink. An extrusion-based 3D bioprinter was used in this work, and astrocytes were extracted from C57Bl/6 mice pups' brain cortices. The bioink was prepared by mixing cortical astrocytes from up to passage 3 to a biomaterial solution composed of gelatin, gelatin-methacryloyl (GelMA), and fibrinogen, supplemented with laminin, which presented optimal bioprinting conditions. The 3D bioprinting conditions minimized cell stress, contributing to the high viability of the astrocytes during the process, in which 74.08% +/- 1.33% of cells were viable right after bioprinting. After 1 week of incubation, the viability of astrocytes significantly increased to 83.54% +/- 3.00%, indicating that the 3D construct represents a suitable microenvironment for cell growth. The biomaterial composition allowed cell attachment and stimulated astrocytic behavior, with cells expressing the specific astrocytes marker glial fibrillary acidic protein (GFAP) and possessing typical astrocytic morphology. This reproducible protocol provides a valuable method to biofabricate 3D neurallike tissue rich in astrocytes that resembles cells' native microenvironment, useful to researchers that aim to understand astrocytes' functionality and their relation to the mechanisms involved in neurological diseases. (AU)

• Processo FAPESP: 18/23039-3 - Bioimpressão 3D de células-tronco neurais visando mimetizar nichos neurogênicos para estudos de doenças neurodegenerativas
• Beneficiário: Bruna Alice Gomes de Melo
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 18/12605-8 - Desenvolvimento de microplataformas brain-on-a-chip para modelagem do sistema nervoso central in vitro
• Beneficiário: Marimélia Aparecida Porcionatto
• Modalidade de apoio: Auxílio à Pesquisa - Temático