Human induced pluripotent stem cell (iPSCs) derived neural models to study brain disease provide an unlimited resource for disease modelling as well as being a tool for drug screening for effective therapies. The limited access to viable patient neurons from brain tissue itself generates the need for functional and reproducible human neuron cell models. Such cells are now increasingly used for drug development studies as well as supporting research into mechanisms and pathways of various neurological diseases.
REPROCELL’s StemRNA™ Neuro cells , when cultured, will form a network of mature neurons that develop increasingly dense synaptic connections over time. These cultures can then be used in an array of assays such as assessment of neurite outgrowth, analysis of cell signalling including electrical action potentials, and gene expression analysis.
Figure 1: Neural stem cells derived from human induced pluripotent stem (iPSCs) cells StemRNA Neuro cells (RCDN001N) are differentiated into neurons using REPROCELL’s supplemented media (Neuro Culture Medium, RCDN101) and coating reagents (Neuro Coat, RCDN201).
Culturing StemRNA Neuro Human iPSCs Neurons
StemRNA Neuro is a single-cell suspension of late stage progenitor cells prepared from disrupted neurospheres. When the cells are plated in 2D culture plate with microwells, cell aggregation is promoted and spheroid formation is induced over a 24-hour culture phase.
Neurite Outgrowth Assays
Neurite outgrowth assays are a simple but efficient quantitative method to investigate compounds that effect neurite formation and toxicity.
Induction of neurite outgrowth is performed by transferring generated neurospheres into plates prepared with Neuro Coat. This promotes attachment of the neurospheres and induces neurite outgrowth over the course of the culture phase. For neurite outgrowth assays, 1-2 neurospheres are plated per well to allow neurites to fully form for high quality immunofluorescent imaging, enabling quantification of neurite outgrowth (Number of Neurites per Neurosphere).
Figure 2. Immunofluorescent analysis of neurite outgrowth using βIII-Tubulin (green) and Hoechst (blue).
Available from REPROCELL
- StemRNA™ Neuro (RCDN001N) — human iPSC neuronal cells.
- Neuro Coat (RCDN201) — an optimized blend of extracellular matrix (ECM) proteins for coating plastic surfaces being used to culture human neurons.
- Neuro Culture Medium (RCDN101) — a culture medium optimized for growth and maturation of StemRNA Neuro cells.
Dr Lyndsey McBroom
Study Director, REPROCELL Europe