Purification of Immature Neuronal Cells from Neural Stem Cell Progeny

• DOI: 10.1371/journal.pone.0020941
• Publication Date: 2011-06-03T20:54:10Z
• Authors: Hassan Azari; Geoffrey W. Osborne; Takahiro Yasuda; Mohammad G. Golmo...; Maryam Rahman; Loic P. Deleyrolle; Ebrahim Esfandiari; David J. Adams; Bjorn Scheffler; Dennis A. Steindler; Brent A. Reynolds
• Keywords: NeuN

Large-scale proliferation and multi-lineage differentiation capabilities make neural stem cells (NSCs) a promising renewable source of for therapeutic applications. However, the practical application neuronal cell replacement is limited by heterogeneity NSC progeny, relatively low yield neurons, predominance astrocytes, poor survival donor following transplantation potential uncontrolled precursor cells. To address these impediments, we have developed method generation highly enriched immature neurons from murine progeny. Adaptation standard procedure in concert with flow cytometry selection, using scattered light positive fluorescent selection based on surface antibody binding, provided near pure (97%) neuron population. Using purified screened panel growth factors found that bone morphogenetic protein-4 (BMP-4) demonstrated strong effect vitro, enhanced their functional maturity. This was maintained into adult mouse striatum where observed 2-fold increase implanted 3-fold NeuN expression. Additionally, neural-colony forming assay (N-CFCA), noted 64 fold reduction bona fide frequency population showed no signs excessive or proliferation. The ability to provide defined populations sources such as may find therapies central nervous system.