Adhesion and Growth of Neuralized Mouse Embryonic Stem Cells on Parylene-C/SiO2 Substrates

Abstract

Neuronal patterning on microfabricated architectures has developed rapidly over the past few years, together with the emergence of soft biocompatible materials and tissue engineering scaffolds. Previously, we introduced a patterning technique based on serum and the biopolymer parylene-C, achieving highly compliant growth of primary neurons and astrocytes on different geometries. Here, we expanded this technique and illustrated that neuralized cells derived from mouse embryonic stem cells (mESCs) followed stripes of variable widths with conformity equal to or higher than that of primary neurons and astrocytes. Our results indicate the presence of undifferentiated mESCs, which also conformed to the underlying patterns to a high degree. This is an exciting and unexpected outcome, as molecular mechanisms governing cell and ECM protein interactions are different in stem cells and primary cells. Our study enables further investigations into the development and electrophysiology of differentiating patterned neural stem cells.

Localisation of oestrogen receptors in stem cells and in stem cell‐derived neurons of the mouse

“…Although the three selected timepoints for neuronal characterization (D7, D14 and D21) do not correspond to any specific embryonic or postnatal developmental stage, in the vast majority of neuronal studies that contain a developmental element, characterization is carried out within this timeframe. 45,[52][53][54] Our data clearly show the presence of all receptors in the mES and in neurons derived from mES. Human ES and EBs express both ERα-66 and ERβ 55 ; however, ERβ is not reliably detectable with currently available antibodies and hence not assayed in this study.…”

“…In our protocol, we obtain NPCs by Day 6 and supplement BDNF and βFGF in the neuronal media to ensure neuronal maturation over the following 3 weeks. Although the three selected timepoints for neuronal characterization (D7, D14 and D21) do not correspond to any specific embryonic or postnatal developmental stage, in the vast majority of neuronal studies that contain a developmental element, characterization is carried out within this timeframe 45,52–54 …”