Department of Otolaryngology, Central Laboratory, Eye Ear Nose and Throat Hospital, Fudan University, Shanghai 200031, China.
The objective of this study is to explore whether olfactory ensheathing cells (OECs) can promote the survival of newborn rat spiral ganglion cells (SGCs) and the underlying possible mechanisms. Co-culture of OECs from adult rats with SGCs from newborn rat cochlea was established and single culture of SGCs acted as control. OECs were obtained and purified based on their special rate of attachment which was different from the other harvested cell types during culture. OECs and SGCs were immunocytochemically characterized and confirmed by expression of low-affinity nerve growth factor receptor p75 or positive label of neuron-specific betaIII-tubulin. To investigate the mechanisms of the role of OECs in survival of SGCs, brain derived neurotrophic factor (BDNF) and anti-BDNF antibody (IgY) were added into the media of the co-cultures respectively, and the surviving SGCs were examined after treatment. Single layer of OECs (92% pure) was seen seven days after plating. Surviving SGCs, which extended their primary neurites, were found on the surface of the layer in the co-cultures. When OECs and SGCs were co-cultured, the number of surviving SGCs was significantly greater than that in the single culture (P<0.01). Nine days after culture, there was even no change in the number of surviving SGCs in the co-culture while the number reduced to almost zero in the single culture. In comparison with co-culture without treatment, addition of BDNF (500 pg/mL) into the media had no obvious promoting effect on the survival of SGCs. The number of surviving SGCs reduced significantly when anti-BDNF antibody was applied into the media of co-cultures (P<0.01). These results suggest that OECs can promote the survival of SGCs when they are co-cultured in vitro. BDNF released from OECs, as one of the survival factors, plays an important role in the survival of SGCs.
PMID: 20401445 [PubMed - in process]