The University of Reading EM Lab (formerly know as CfAM) has been assisting in our studies of 3D human neural cell cultures derived from a type of neural stem cells (normal human neural progenitors – NHNP). Here at Reading I’ve been investigating the electrophysiological properties of cultures of neurons and glial cells differentiated from NHNP, grown in a 3D substrate called Alvetex. In order to see how cells are expanding and morphologically orientate themselves in Alvetex, I’ve been using immunofluorescence and SEM. Not only have the EM Lab done the hard graft of the SEM work, one of our images went up in their image of the month hall of fame for 2013. All part of the art of electrophysiology!
SEM (left) and immunofluorescent staining (right) NHNP cells grown in 3D scaffold SEM imaging: Matthew Spink @ EMLab, Immunostaining: Immy Smith @ UoR ERG. Cells – Lonza NHNP, Scaffold – Alvetex by Reinnervate, Antibodies – B-tubulin III (Millipore) in green, GFAP (Dako) in red, hoechst 33342 counterstain and secondary antibodies from Life Technologies)
This scanning electron micrograph shows a normal human neural progenitor cell (supplied by Lonza) which has been false coloured in blue. A normal human neural progenitor cell is a type of stem cell which forms neurons and glia in the brain during development. This cell has not yet commenced differentiating into neural lineages, and is still in the stem cell stage. The cell is grown in a 3D culture scaffold called Alvetex. This work is being undertaken by Dr Immy Smith who is part of the Electrophysiology Research Group in the School of Pharmacy. The work forms part of a project investigating functional differences in these cells grown in 2D vs. 3D to produce models for neurotoxicity and pharmacological studies. Sample preparation and microscopy assistance was provided by Matthew Spink.