This cell population could be enriched greater than 1.5-fold (74-76%) when a mechanical isolation step was introduced, similar to the results of Reubinoff et al. and Zhang et al., who reported that highly enriched (>95%) hES-derived NPCs could be obtained by isolation of tubular structures from hES cell cultures or EB outgrowths. Thus, a combination of serum-free conditions and mechanical isolation is an efficient approach for deriving populations of highly enriched NPCs or putative neurons from primate ES cells. Viagra super active Canadian pharmacy
Representatives of the two nervous system lineages, neurons and astrocytes, were present in NPC cultures. The third lineage, oligodendrocytes, was not evaluated here but has been reported at low frequency in hES cell progeny. We made little effort to define nonneural phenotypes, despite the recognition that nestin-positive cells also represent the starting point for derivation of pancreatic islet phenotypes.
Differences in the differentiation of monkey and mouse ES cells noted here may suggest suboptimal conditions in the neuronal induction medium or reflect a higher degree of heterogeneity in monkey ES cell populations. Thus, monkey EBs after expansion may contain fewer NPC progenitor cells. The ICC analyses clearly demonstrated that complex/cystic, but not early, monkey EBs (Day 15 EBs) contained cells expressing antigenetic markers representative of all three germ layers.