3D Astrogliosis Model with bFGF and GFAP Expression Profiles Corresponding to an MCAO-injured Brain

• Published in: Biotechnology and bioprocess engineering Vol. 23; no. 5; pp. 588 - 597
• Main Authors: Min, Seul Ki, Shim, Hyeon Jung, Shin, Hwa Sung
• Format: Journal Article
• Language: English
• Published: Seoul The Korean Society for Biotechnology and Bioengineering 01.09.2018; Springer Nature B.V; 한국생물공학회
• Subjects: Adenosine; animal experimentation; Animal research; animal use replacement; animals; Astrocytes; Biotechnology; brain; Brain damage; Brain diseases; Brain injury; Cell culture; Cerebral blood flow; Chemistry; Chemistry and Materials Science; Confocal microscopy; Drug development; drugs; ethics; Expenditures; Fibroblast growth factor 2; Glial fibrillary acidic protein; Gliosis; in vitro culture; in vitro studies; Industrial and Production Engineering; Ischemia; Nanofibers; Neuroprotection; Occlusion; quantitative polymerase chain reaction; Reperfusion; Research Paper; reverse transcriptase polymerase chain reaction; screening; Signaling; Stroke; Three dimensional models; transforming growth factor beta 1; Transforming growth factor-b1; Western blotting; 생물공학; MCAO; TGF-β1; astrogliosis; 3D nanofiber culture model; bFGF; GFAP
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-018-0207-7

Various brain diseases including ischemic stroke cause profound changes in the cells of the brain. In order to develop neuroprotective drugs useful in overcoming brain diseases, pathologic cellular changes such as astrogliosis in lesion sites have to be better understood. Many drug development trials, which are often unsuccessful, require the sacrifice of large numbers of animals, leading to ethical issues and enormous expenditures of time and financial resources. Therefore, initial screening should be performed using in vitro models that are similar to the in vivo context in order to mitigate these concerns as much as possible. In this study, we developed an astrocyte culture model employing a three-dimensional (3D) nanofibrous mat that may be able to replace vast numbers of animal experiments in studies of middle cerebral artery occlusion (MCAO), ischemic stroke and reperfusion, and related brain damage. To induce artificial activation of astrocytes, 3D cultures were exogenously treated with TGF-β1. Subsequent changes in astrocytes were monitored and characterized by using qRT-PCR, western blot, and confocal microscopy of immunocytochemically stained cells. We observed that, upon treatment with TGF-β1, bFGF was upregulated and accompanied a change in GFAP expression in 3D astrocyte cultures, similar to that observed in MCAO-injured brain tissue. These changes were demonstrated to be associated with bFGF signaling through the application of 5′-Deoxy- 5′-(methylthio)adenosine treatment. We conclude that astrocytes cultured with TGF-β1 in our 3D nanofiber culture model mimics in vivo -like bFGF signaling and could be used as an in vitro model system to study astrogliosis after MCAO and reperfusion.