Abstract
A novel in vitro model of glial scarring was developed by adapting a primary cell-based system previously used for studying neuroinflammatory processes in neurodegenerative disease. Midbrains from embryonic day 14 Fischer 344 rats were mechanically dissociated and grown on poly-d-lysine coated 24 well plates to a confluent layer of neurons, astrocytes, and microglia. The culture was injured with either a mechanical scrape or foreign-body placement (segments of 50 μm diameter stainless steel microwire), fixed at time points from 6 h to 10 days, and assessed by immunocytochemistry. Microglia invaded the scraped wound area at early time points and hypertrophied activated astrocytes repopulated the wound after 7 days. The chronic presence of microwire resulted in a glial scar forming at 10 days, with microglia forming an inner layer of cells coating the microwire, while astrocytes surrounded the microglial core with a network of cellular processes containing upregulated GFAP. Vimentin expressing cells and processes were present in the scrape at early times and within the astrocyte processes forming the glial scar. Neurons within the culture did not repopulate the scrape wound and did not respond to the microwire, although they were determined to be electrically active through patch clamp recording. The time course and relative positions of the glia in response to the different injury paradigms correlated well with stereotypical in vivo responses and warrant further work in the development of a functional in vitro test bed.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 5368-5376 |
| Number of pages | 9 |
| Journal | Biomaterials |
| Volume | 27 |
| Issue number | 31 |
| DOIs | |
| State | Published - Nov 2006 |
| Externally published | Yes |
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Keywords
- Biocompatibility
- Brain
- Cell culture
- Foreign-body response
- In vitro test
- Neural prosthesis
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Biomedical Engineering
Cite this
In vitro model of glial scarring around neuroelectrodes chronically implanted in the CNS. / Polikov, Vadim S.; Block, Michelle L.; Fellous, Jean-Marc; Hong, Jau Shyong; Reichert, W. Monty.
In: Biomaterials, Vol. 27, No. 31, 11.2006, p. 5368-5376.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - In vitro model of glial scarring around neuroelectrodes chronically implanted in the CNS
AU - Polikov, Vadim S.
AU - Block, Michelle L.
AU - Fellous, Jean-Marc
AU - Hong, Jau Shyong
AU - Reichert, W. Monty
PY - 2006/11
Y1 - 2006/11
N2 - A novel in vitro model of glial scarring was developed by adapting a primary cell-based system previously used for studying neuroinflammatory processes in neurodegenerative disease. Midbrains from embryonic day 14 Fischer 344 rats were mechanically dissociated and grown on poly-d-lysine coated 24 well plates to a confluent layer of neurons, astrocytes, and microglia. The culture was injured with either a mechanical scrape or foreign-body placement (segments of 50 μm diameter stainless steel microwire), fixed at time points from 6 h to 10 days, and assessed by immunocytochemistry. Microglia invaded the scraped wound area at early time points and hypertrophied activated astrocytes repopulated the wound after 7 days. The chronic presence of microwire resulted in a glial scar forming at 10 days, with microglia forming an inner layer of cells coating the microwire, while astrocytes surrounded the microglial core with a network of cellular processes containing upregulated GFAP. Vimentin expressing cells and processes were present in the scrape at early times and within the astrocyte processes forming the glial scar. Neurons within the culture did not repopulate the scrape wound and did not respond to the microwire, although they were determined to be electrically active through patch clamp recording. The time course and relative positions of the glia in response to the different injury paradigms correlated well with stereotypical in vivo responses and warrant further work in the development of a functional in vitro test bed.
AB - A novel in vitro model of glial scarring was developed by adapting a primary cell-based system previously used for studying neuroinflammatory processes in neurodegenerative disease. Midbrains from embryonic day 14 Fischer 344 rats were mechanically dissociated and grown on poly-d-lysine coated 24 well plates to a confluent layer of neurons, astrocytes, and microglia. The culture was injured with either a mechanical scrape or foreign-body placement (segments of 50 μm diameter stainless steel microwire), fixed at time points from 6 h to 10 days, and assessed by immunocytochemistry. Microglia invaded the scraped wound area at early time points and hypertrophied activated astrocytes repopulated the wound after 7 days. The chronic presence of microwire resulted in a glial scar forming at 10 days, with microglia forming an inner layer of cells coating the microwire, while astrocytes surrounded the microglial core with a network of cellular processes containing upregulated GFAP. Vimentin expressing cells and processes were present in the scrape at early times and within the astrocyte processes forming the glial scar. Neurons within the culture did not repopulate the scrape wound and did not respond to the microwire, although they were determined to be electrically active through patch clamp recording. The time course and relative positions of the glia in response to the different injury paradigms correlated well with stereotypical in vivo responses and warrant further work in the development of a functional in vitro test bed.
KW - Biocompatibility
KW - Brain
KW - Cell culture
KW - Foreign-body response
KW - In vitro test
KW - Neural prosthesis
UR - http://www.scopus.com/inward/record.url?scp=33746255986&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33746255986&partnerID=8YFLogxK
U2 - 10.1016/j.biomaterials.2006.06.018
DO - 10.1016/j.biomaterials.2006.06.018
M3 - Article
VL - 27
SP - 5368
EP - 5376
JO - Biomaterials
JF - Biomaterials
SN - 0142-9612
IS - 31
ER -