Workflows for Culturing Primary-Derived Neural Stem and Progenitor Cells

Neural stem and progenitor cells (NSCs) have traditionally been cultured using the neurosphere system. However, the adherent monolayer method has recently become more popular and may be preferable to the neurosphere assay when performing certain techniques (e.g. transfection1). It has been reported that NSCs cultured under adherent conditions in the presence of extracellular matrices undergo symmetrical divisions in long term cultures maintaining a more "pure" population of stem cells, as opposed to the heterogeneous population of cells present in neurospheres.2-4

Both culture systems are useful; you will need to determine which one is more applicable to the questions being addressed by your research. Please refer to the table below for a comparison of the two methods:

Neurosphere Adherent Monolayer
Heterogeneous population Claimed to be less heterogeneous2-4
Cell aggregates in suspension Cell monolayer attached to substrate
No substrate required Substrate required (PDL, Laminin)
Owing to 3D aggregate structure, cells in center of neurosphere may be deprived of nutrients 2D monolayer format allows for uniform exposure to culture media and nutrients
Neurosphere 3D structures thought to be more reminiscent of in vivo neurogenic niche 2D monolayer system created when cells exposed to extracellular matrix and cultured on a flat plastic dish
May promote more glial cell differentiation in long term cultures Claimed to promote less glial cell differentiation in long term cultures
Neurospheres are dissociated by various methods (trituration, enzymatic, chemical) Monolayers are detached with enzymes (trypsin or ACCUTASE™)

Although our NeuroCult™ proliferation media were originally developed to support the proliferation of NSCs in neurosphere suspension cultures, they are also fully compatible with adherent culture systems. This is important and also convenient, because it allows for the direct comparison of the functional properties of NSCs cultured in different culture systems using a controlled medium formulation. Both culture systems are capable of supporting the isolation of a population cells capable of self-renewal and multi-lineage differentiation. A comparison of these two methods for culturing mouse NSCs in NeuroCult™ medium is shown in this poster (2010 SFN Meeting).

Please refer to our website for more information on our mouse, rat and human NeuroCult™ proliferation media, including detailed protocols for the expansion of NSCs in both neurosphere and adherent monolayer cultures (see Manuals, under Technical Resources tab):
For more information, please refer to the MINI-REVIEW: Neural Stem Cells or contact

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