Production and Maintenance of Two Populations of Neural Progenitor Cells with Rostral and Caudal Properties from Human Embryonic Stem Cells (Pages: 0-0)

Nemati Sh *, Hatami M , Hemmesi K , Kiani S , Baharvand H ,


Objective: Availability of human embryonic stem cells (hESC) has enhanced human neural differentiation research. While differentiation is directed towards the neural lineage, lack of an optimal protocol implies generation of other lineage cells as contaminants. Thus, one major challenge in the field is to generate a homogeneous and renewable, easy to culture, neural progenitor (NP) cell population committed to the neural lineage, capable of serving as an unlimited lineage-restricted cell source for replacement therapy and novel drug screening and/or for other studies. Materials and Methods: Here, we present two pure populations of long-term self-renewing rosette-type hESC-derived neural progenitor cells (hES-NP cells) with Rostral and Caudal properties induced by retinoic acid and noggin(RA+ population & RA- population), which exhibit extensive self-renewal, clonogenicity, and stable neurogenesis Results: Flowcytometry and immunocytochemistry analysis of both populations showed increase in expression of Nestin, SOX-1, and Pax-6.Then real-time PCR analysis showed increase expression of Hoxc5 and Otx2, in RA+ population rather than RA- population. In addition, hES-NP cells maintained their developmental potential through long-term storage in liquid nitrogen and multiple freeze–thaw cycles. Conclusion: These results demonstrate that hES-NP cells have the ability to provide an expandable and unlimited human cell source that can develop into specific neuronal and glial subtypes.