Abstract
This chapter describes protocols to establish simplified in vitro assays of Schwann cell (SC) differentiation in the absence of neurons. The assays are based on the capacity of isolated primary SCs to increase or decrease the expression of myelination-associated genes in response to the presence or absence of cell permeable analogs of cyclic adenosine monophosphate (cAMP). No special conditions of media or substrates beyond the administration or removal of cAMP analogs are required to obtain a synchronous response on differentiation and dedifferentiation. The assays are cost-effective and far easier to implement than traditional myelinating SC-neuron cultures. They are scalable to a variety of plate formats suited for downstream experimentation and analysis. These cell-based assays can be used as drug discovery platforms for the evaluation of novel agents controlling the onset, maintenance, and reversal of the differentiated state using any typical adherent SC population.
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 213-232 |
| Number of pages | 20 |
| DOIs | |
| State | Published - Jan 1 2018 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1739 |
| ISSN (Print) | 1064-3745 |
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Keywords
- c-Jun
- cAMP
- Cell morphology
- In vitro assays
- Krox-20
- MAG
- Myelin markers
- O1
- Primary cultures
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Scalable differentiation and dedifferentiation assays using neuron-free schwann cell cultures. / Monje, Paula V.
Methods in Molecular Biology. Humana Press Inc., 2018. p. 213-232 (Methods in Molecular Biology; Vol. 1739).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Scalable differentiation and dedifferentiation assays using neuron-free schwann cell cultures
AU - Monje, Paula V
PY - 2018/1/1
Y1 - 2018/1/1
N2 - This chapter describes protocols to establish simplified in vitro assays of Schwann cell (SC) differentiation in the absence of neurons. The assays are based on the capacity of isolated primary SCs to increase or decrease the expression of myelination-associated genes in response to the presence or absence of cell permeable analogs of cyclic adenosine monophosphate (cAMP). No special conditions of media or substrates beyond the administration or removal of cAMP analogs are required to obtain a synchronous response on differentiation and dedifferentiation. The assays are cost-effective and far easier to implement than traditional myelinating SC-neuron cultures. They are scalable to a variety of plate formats suited for downstream experimentation and analysis. These cell-based assays can be used as drug discovery platforms for the evaluation of novel agents controlling the onset, maintenance, and reversal of the differentiated state using any typical adherent SC population.
AB - This chapter describes protocols to establish simplified in vitro assays of Schwann cell (SC) differentiation in the absence of neurons. The assays are based on the capacity of isolated primary SCs to increase or decrease the expression of myelination-associated genes in response to the presence or absence of cell permeable analogs of cyclic adenosine monophosphate (cAMP). No special conditions of media or substrates beyond the administration or removal of cAMP analogs are required to obtain a synchronous response on differentiation and dedifferentiation. The assays are cost-effective and far easier to implement than traditional myelinating SC-neuron cultures. They are scalable to a variety of plate formats suited for downstream experimentation and analysis. These cell-based assays can be used as drug discovery platforms for the evaluation of novel agents controlling the onset, maintenance, and reversal of the differentiated state using any typical adherent SC population.
KW - c-Jun
KW - cAMP
KW - Cell morphology
KW - In vitro assays
KW - Krox-20
KW - MAG
KW - Myelin markers
KW - O1
KW - Primary cultures
UR - http://www.scopus.com/inward/record.url?scp=85044221921&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85044221921&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7649-2_14
DO - 10.1007/978-1-4939-7649-2_14
M3 - Chapter
T3 - Methods in Molecular Biology
SP - 213
EP - 232
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -