Enhanced Yield of Neuroepithelial Precursors and Midbrain-Like Dopaminergic Neurons from Human Embryonic Stem Cells Using the Bone Morphogenic Protein Antagonist Noggin

Abstract

STEM CELLS 2007;25:411-418

Differentiated Parkinson patient-derived induced pluripotent stem cells grow in the adult rodent brain and reduce motor asymmetry in Parkinsonian rats

“…S1) using the stromal feeder cell-based differentiation protocol (14,18). Consistent with our previously published findings from other PDiPS cell lines (7), we did not observe major differences in DA differentiation when comparing the factor-carrying and the factor-free PDiPS cells with hES cells or non-PDiPS cells that had been derived from a subject who did not have PD (Fig.…”

“…However, we and others have previously shown that tumor formation can occur when transplanting human pluripotent stem cellderived cells (14,15), demonstrating a requirement to further purify cells before transplantation. Therefore, we performed FACS of differentiated PDiPS cells for NCAM before transplantation and did not observe signs of tumor formation in the grafts.…”

“…Therefore, we believe the PDiPS cells have most likely been inefficiently patterned toward an A9 DA neuronal phenotype during in vitro differentiation. Notably, hES cellderived DA neurons also have not yet been shown to coexpress TH and foxA2 after transplantation using different differentiation protocols, including ours (12)(13)(14)(15), indicating that new strategies toward phenotypic patterning of hES and hiPSs cells during in vitro differentiation have to be pursued. During the course of this study, however, we and others have developed protocols that generate foxA2 + floor plate cells from hES cells (28) or foxA2 + DA neurons from hES and PDiPS cells in vitro (29), which provide encouraging cell sources for future transplantation studies to further improve axonal outgrowth and behavioral outcomes.…”

“…Human stem cells were maintained on mitomycin C-inactivated human D551 fibroblasts (American Type Culture Collection) and were differentiated according to a stromal feeder cell-based protocol (18), which was modified through the addition of noggin (300 ng/mL) during the first 21 d of differentiation to improve neuroectodermal differentiation (14).…”

“…Several studies have shown that some patients with PD benefit from the transplantation of human fetal cells (10,11), but limited tissue availability requires alternative cellular sources. Human ES (hES) cells have been transplanted into animal models of PD after in vitro differentiation into neural precursors (12) or DA neurons (13,14), and partial functional recovery was observed in some of these reports (12,13). One study reported complete functional recovery after transplantation of differentiated hES cells but severe graft overgrowth was found in engrafted animals (15), emphasizing efforts to purify hES cellderived neurons via FACS before transplantation (16,17).…”