ABSTRACT
New small molecules that regulate the step-wise differentiation of human pluripotent stem cells into dopaminergic neurons have been identified. The steroid, guggulsterone, was found to be the most effective inducer of neural stem cells into dopaminergic neurons. These neurons are extensively characterized and shown to be functional. We believe this new approach offers a practical route to creating neurons of sufficient quality to be used to treat Parkinson's disease patients.
Subject(s)
Cell Differentiation/drug effects , Dopaminergic Neurons/cytology , Neural Stem Cells/cytology , Pluripotent Stem Cells/cytology , Pregnenediones/pharmacology , Cell Culture Techniques , Cell Line , Cell Transplantation/methods , Dopamine/metabolism , Dopaminergic Neurons/metabolism , Dopaminergic Neurons/physiology , Gene Expression Profiling , Humans , Membrane Potentials/drug effects , Neural Stem Cells/metabolism , Neural Stem Cells/physiology , Oligonucleotide Array Sequence Analysis , Parkinson Disease/surgery , Patch-Clamp Techniques , Pluripotent Stem Cells/metabolism , Pluripotent Stem Cells/physiology , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Human parthenogenetic stem cells are derived from the inner cell mass of blastocysts obtained from unfertilized oocytes that have been stimulated to develop without any participation of male gamete. As parthenogenesis does not involve the destruction of a viable human embryo, the derivation and use of human parthenogenetic stem cells does not raise the same ethical concerns as conventional embryonic stem cells. Human parthenogenetic stem cells are similar to embryonic stem cells in their proliferation and multilineage in vitro differentiation capacity. The aim of this study is to derive multipotent neural stem cells from human parthenogenetic stem cells that are stable to passaging and cryopreservation, and have the ability to further differentiate into functional neurons. Immunocytochemistry, quantitative real-time PCR, or FACS were used to confirm that the derived neural stem cells express neural markers such as NES, SOX2 and MS1. The derived neural stem cells keep uniform morphology for at least 30 passages and can be spontaneously differentiated into cells with neuron morphology that express TUBB3 and MAP2, and fire action potentials. These results suggest that parthenogenetic stem cells are a very promising and potentially unlimited source for the derivation of multipotent neural stem cells that can be used for therapeutic applications.
