Rat Neural Stem cells (rat NSCs) can be maintained in vitro as self renewing multi-potent cells that can be induced to differentiate into astrocytes, neurons, or oligodendrocytes by manipulating culture conditions (1). Therefore, Rat NSCs have the potential of providing an unlimited supply of cells to study differentiation of NSCs to neuronal cell types. Due to the host of neurodegenerative diseases a vast amount of research is generated to study the promise of potential therapeutics involving NSCs.
Currently, the most utilized method to determine the undifferentiated state of Rat Neural Stem Cells is to analyze the expression of Sox2 and Nestin (3,4). Neural progenitor cells expressing both Sox2 and Nestin have been shown to self-replicate as well as give rise to astrocytes, neurons, and oligodendrocytes under the correct culture conditions (3, 4). Beta-III-Tubulin is a specific component of microtubules found in neuronal axons and cell bodies (5). Upon differentiation to a neuronal lineage, Sox2 and Nestin expression levels decrease while Beta-III-Tubulin expression increases.
The FlowCellect™ Rat Neural Stem Cell Characterization kit for neurons is designed for cell biology applications. The kit uses an antibody detection method, utilizing fluorescently conjugated primary antibodies to detect Sox2, Nestin and Beta-III-Tubulin. The FlowCellect™ Rodent Neural Stem Cell Characterization Kit for neurons is optimized for flow cytometers with a single blue (488 nm) laser light source.
The kit is optimized for use on the Guava EasyCyte™ Plus cytometer that requires cell concentrations to be less than 500 cells per microliter. The kit does not include culture media or other materials that should be readily available in a common laboratory setting that routinely performs experiments of the type described here.