In Vitro Pancreatic Neuroplasticity Assay: Studying Changes in GI Tract Neurons Exposed to Healthy and Cancerous Human Pancreatic Tissue Extracts

- The dorsal root ganglion or DRG are sensory neurons that relay messages from the periphery to the central nervous system. In contrast, myenteric plexus neurons are involved in peristaltic movements of the bowels.

In cancer conditions such as pancreatic cancer, these neurons alter morphologically and functionally. To study the neuronal morphology changes, seed the neural cells on coverslips pre-coated with laminin protein, placed in a multiwell plate, and top the cells with the neural basal medium.

Laminin protein promotes neurite outgrowth. Incubate the cells overnight. Next, add the pancreatic cancer tissue extract media to the first three wells and healthy pancreatic tissue extract media to the subsequent three wells. Incubate the plate. Now, perform immunohistochemical staining using neuron-specific antibody markers, which bind to neurons. Place the coverslip on the glass slide and place the slide under an inverted microscope. Measure neurite density using the software.

The DRG neurons cultured in pancreatic cancer tissue extract usually show greater neurite density than in healthy pancreatic tissue extract. In contrast, myenteric plexus neurons form longer neurites than in he