RESUMEN

Schwann cells develop from multipotent neural crest stem cells and are important for neuronal survival, maintenance of axonal integrity, and myelination. We used transgenic mice expressing green fluorescent protein in a tissue-specific manner to isolate viable, pure populations of neural crest stem cells and developing Schwann cells, which are not readily accessible by microdissection. Starting with the minute amounts of RNA obtained, a two-round amplification procedure was used to achieve reproducible DNA array hybridizations. We validated our screening procedure by comparisons with the literature and by in situ hybridization. Stage-to-stage comparisons and hierarchical clustering for neural crest and five stages of Schwann cell development suggest a wealth of candidates for genes involved in stem cell regulation and in early Schwann cell development. The combination of methods applied in this study should be generally useful for isolating and profiling other stem cell and difficult to isolate cell populations.

Asunto(s)

Perfilación de la Expresión Génica/métodos , Cresta Neural/citología , Células de Schwann/citología , Células de Schwann/metabolismo , Células Madre/citología , Células Madre/metabolismo , Animales , Recuento de Células , Diferenciación Celular/fisiología , Linaje de la Célula/genética , Separación Celular/métodos , Células Cultivadas , Citometría de Flujo , Genes Reporteros , Edad Gestacional , Proteínas Fluorescentes Verdes , Hibridación in Situ , Proteínas Luminiscentes/biosíntesis , Proteínas Luminiscentes/genética , Ratones , Ratones Transgénicos , Análisis de Secuencia por Matrices de Oligonucleótidos , Valor Predictivo de las Pruebas , ARN/química , ARN/aislamiento & purificación , Reproducibilidad de los Resultados