Co-culture of neuroepithelial stem cells with interstitial cells of Cajal results in neuron differentiation.

OBJECTIVE

The interstitial cells of Cajal (ICCs) interact morphologically and functionally with the elements of the enteric nervous system in the digestive tract. However, direct evidence that ICCs participate in the differentiation of the enteric nervous system is lacking. In this work, we examined in co-culture experiments whether ICCs could stimulate the differentiation of neuroepithelial stem cells (NESCs) to neurons.

METHODS

NESCs were harvested from the neural tube of embryonic (E11.5) rats, and ICCs were isolated from the colons of newborn rats. Various cell types were identified immunohistochemically.

RESULTS

NESCs reacted with antibodies to the stem-cell marker nest in; ICCs reacted with c-kit antibodies. NESCs, when differentiated into astrocytes, were identified with a marker GFAP, and neurons with marker MAP2. NESCs co-cultured with ICCs, compared with NESCs cultured alone, yielded a significantly greater number of cells positive for the neuronal markers PGP9.5 and nNOS. The co-cultured NESCs also produced more PGP9.5 and nNOS proteins, as measured by Western blotting. In addition, co-cultured ICCs connected morphologically with differentiated NESCs.

CONCLUSION

These in vitro findings demonstrated that ICCs could induce the neuronal differentiation of NESCs, which connected with differentiated neurons into a network morphologically. The findings provide an experimental basis for in vivo application of the simultaneous transplantation of NESCs and ICCs.