Spinal motor neurons from neonatal rats：purification, culture and identification / 中国组织工程研究

ABSTRACT

BACKGROUND:neurons are post-mitotic cels that are difficult to survive. Isolation, purification and culture of spinal motor neurons are technical difficulties of cel culture technology. OBJECTIVE:To establish the culture system of spinal motor neurons from neonatal rats and to identify and determine the purity of spinal cord neurons. METHODS: Ventral spinal cord tissues from neonatal rats were made into cel suspension that was subjected to density gradient centrifugation and differential adherence folowed by purification culture. Then, the cels on cover plates were identified and classified using immunocytochemical double staining method, and the content of cel components was calculated in combination with fluorescent Hoechst nuclear staining. RESULTS AND CONCLUSION:The cels adhered wel, and the neurons accounted for 85.8%, among which, motor neurons reached 71.6%, astrocytes accounted for 7.8%, cels negative for neurofilament 200 and glial fibrilary acidic protein were 6.4%. These findings indicate that the ventral spinal cord tissues from neonatal rats combined with density gradient centrifugation and differential adherence can develop high-purity spinal motor neurons.