3D Bioprinting of Murine Cortical Astrocytes for E... - BV FAPESP
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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
de Melo, Bruna A. G. [1] ; Cruz, Elisa M. [1] ; Ribeiro, Tais N. [1] ; Mundim, V, Mayara ; Porcionatto, Marimelia A. [2]
Total Authors: 5
Affiliation:
[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
Total Affiliations: 2
Document type: Journal article
Source: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS; n. 173 JUL 2021.
Web of Science Citations: 0
Abstract

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)

FAPESP's process: 18/23039-3 - 3D Bioprinting of neural stem cells aiming the fabrication of neurogenic niches for the study of neurodegenerative diseases treatment
Grantee:Bruna Alice Gomes de Melo
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 18/12605-8 - Development of brain-on-a-chip microplataforms for in vitro modeling of the central nervous system
Grantee:Marimélia Aparecida Porcionatto
Support Opportunities: Research Projects - Thematic Grants