Phenotype-based primary screening for drugs promoting neuronal subtype differentiation in embryonic stem cells with light microscope / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 373-380, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-336781
ABSTRACT
<p><b>OBJECTIVE</b>To set up a platform for phenotype-based primary screening of drug candidates promoting neuronal subtype differentiation in embryonic stem cells (ES) with light microscope.</p><p><b>METHODS</b>Hanging drop culture 4-/4+ method was employed to harvest the cells around embryoid body (EB) at differentiation endpoint. Morphological evaluation for neuron-like cells was performed with light microscope. Axons for more than three times of the length of the cell body were considered as neuron-like cells. The compound(s) that promote neuron-like cells was further evaluated. Icariin (ICA, 10(-6)mol/L) and Isobavachin (IBA, 10(-7)mol/L) were selected to screen the differentiation-promoting activity on ES cells. Immunofluorescence staining with specific antibodies (ChAT, GABA) was used to evaluate the neuron subtypes.</p><p><b>RESULTS</b>The cells treated with IBA showed neuron-like phenotype, but the cells treated with ICA did not exhibit the morphological changes. ES cells treated with IBA was further confirmed to be cholinergic and GABAergic neurons.</p><p><b>CONCLUSION</b>Phenotypic screening with light microscope for molecules promoting neuronal differentiation is an effective method with advantages of less labor and material consuming and time saving, and false-positive results derived from immunofluorescence can be avoided. The method confirms that IBA is able to facilitate ES cells differentiating into neuronal cells, including cholinergic neurons and GABAergic neurons.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Phenotype
/
Physiology
/
Cell Differentiation
/
Cell Line
/
Cell Biology
/
Drug Evaluation, Preclinical
/
Embryonic Stem Cells
/
Embryoid Bodies
/
Methods
/
Nerve Regeneration
Type of study:
Diagnostic study
/
Screening study
Limits:
Animals
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2012
Type:
Article
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