Development of a novel method to purify and culture mature rat astrocytes
Abstract/Contents
- Abstract
- Astrocytes constitute more than half of the human brain, yet their functions have yet to be fully elucidated largely due to a lack of a good and pure in vitro culture system. The inability to purify and culture brain astrocytes has long hindered studies of their function. Although astrocyte progenitor cells can be cultured from neonatal brain, culture of more differentiated mature astrocytes from postnatal brain has not previously been possible. Here I report a new method for the prospective purification of astrocytes by immunopanning (IP-astrocytes) and have used this method to investigate several fundamental properties, functions of astrocytes and neuron-glial interactions. I show that these highly purified astrocytes, like neurons, undergo apoptosis in culture, but that vascular cells as well as exogenous HBEGF and Wnt7a strongly promote their survival in serum-free culture. Similarly, I found that developing astrocytes in vivo undergo apoptosis and that the vast majority of surviving astrocytes in vivo contact blood vessels and hypothesise that developing astrocytes may normally be matched to blood vessels by competing for a limited supply of vascular trophic factors. In contrast to the more traditional neonatal astrocyte cultures prepared via a method developed by McCarthy and de Vellis (1980) (MD-astrocytes), the gene profiles and functions of the purified postnatal astrocytes in serum-free culture much more closely resembled that of astrocytes in vivo. These astrocytes also strongly promoted synapse formation and function. I have also identified genes that are induced by serum and show that serum exposure to astrocytes causes massive and irreversible changes of gene expression as well as astrocyte function. In addition, I have highlighted several features of MD-astrocytes that are different from IP-astrocytes. Together, the data suggests that I have developed a culture system that is more representative of the in vivo state. This culture system will be of interest to a broad range of neuroscientists as it provides a simplified and tractable system to study the functions and properties of astrocytes both in the normal and diseased states.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2011 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Foo, Lynette Caizhen | |
|---|---|---|
| Associated with | Stanford University, Neurosciences Program | |
| Primary advisor | Barres, Ben | |
| Thesis advisor | Barres, Ben | |
| Thesis advisor | McConnell, Susan K | |
| Thesis advisor | Shen, Kang, 1972- | |
| Thesis advisor | Talbot, William | |
| Advisor | McConnell, Susan K | |
| Advisor | Shen, Kang, 1972- | |
| Advisor | Talbot, William |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Lynette Caizhen Foo. |
|---|---|
| Note | Submitted to the Program in Neurosciences. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2011. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2011 by Lynette Caizhen Foo
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