Isolation and characterization of duck embryonic neural stem and progenitor cells

Abstract

Both embryonic and adult neural stem cells (NSC) of rodents and humans have been isolated and cultured in vitro to date, and they are thought to have tremendous clinical promise in restoring the diseased or injured central nervous system. However, there are few counterpart reports on neural stem cells from birds. This study explored the isolation and culture system of duck neural stem and progenitor cells (NSPC) and investigated their major biological properties. Cells from the dorsal ventricular ridges of 10- to 13-d embryos were isolated, cultured, and purified by using a neurosphere assay. Growth kinetics and karyotype were analyzed. The differentiation potential of NSPC was detected by immunofluorescence. Apoptosis and acetylation level of histone 4 lysine 12 (H4K12) were assessed. Results indicated that the nestin-positive neurospheres derived from duck embryos were able to self-renew and differentiate into neurons, astrocytes, and oligodendrocytes, and were prone to be transfected with exogenous genes. Karyotype analysis showed that 95% (38 out of 40) of cells of the population were diploid. Apoptosis detection indicated that the apoptotic rate was elevated with increasing passage number and culture time. The cells were highly acetylated and exhibited typical NSPC properties. Efficiently transfected with fluorescent genes, they were available for gene therapy and suitable for research on intracellular distribution of proteins of interest.