Researchers have differentiated hESCs into forebrain, midbrain and hindbrain. However, observing cell interpretation of signals to form dorsal-ventral patterning tissues with sonic hedgehog (SHH) gene is necessary. We used micropatterning within different timeframes to determine how cell fates shift across the dorsal-ventral axis. This reveals how sensitive the cells are to this signal and how early they should read it to become ventral. hESCs were routinely fed initially to remain in its pluripotent state. After cell seeding, researchers treated cells with three different mediums: N1 control, containing N2B27, RI, and Penicillin-Streptomycin, N2, containing N1, LDN dilution to inhibit BMP, and SB to inhibit TGFβ1, and N3, containing N2 and CHIR dilution. Researchers separated the cells into an IBIDI plate and treated each well with one of the three reagents. Throughout experimentation, researchers added varying concentrations of CHIR and FGF8 to experimental IBIDI wells. After cell fixation, micro-patterning was analyzed.

Rice University,  National Science Foundation (NSF) award #EEC-1406885

Researchers should cite this work as follows:

• Rubiona Grainger, Rice University, NEWT Center (2022), "Your Smart Brain," https://nanohub.org/resources/36327.

  BibTex | EndNote

1. brain
2. cells
3. hESCs
4. micropatterning
5. sonic hedgehog (SHH) gene