Xeno-Free Defined Conditions for Culture of Human Embryonic Stem Cells, Neural Stem Cells and Dopaminergic Neurons Derived from Them
نویسندگان
چکیده
BACKGROUND Human embryonic stem cells (hESCs) may provide an invaluable resource for regenerative medicine. To move hESCs towards the clinic it is important that cells with therapeutic potential be reproducibly generated under completely defined conditions. METHODOLOGY/PRINCIPAL FINDINGS Here we report a four-step scalable process that is readily transferable to a Good Manufacture Practice (GMP) facility for the production of functional dopaminergic neurons from hESCs for potential clinical uses. We show that each of the steps (propagation of ESC-->generation of neural stem cells (NSC)-->induction of dopaminergic precursors-->maturation of dopaminergic neurons) could utilize xeno-free defined media and substrate, and that cells could be stored at intermediate stages in the process without losing their functional ability. Neurons generated by this process expressed midbrain and A9 dopaminergic markers and could be transplanted at an appropriate time point in development to survive after transplant. CONCLUSIONS/SIGNIFICANCE hESCs and NSCs can be maintained in xeno-free defined media for a prolonged period of time while retaining their ability to differentiate into authentic dopaminergic neurons. Our defined medium system provides a path to a scalable GMP-applicable process of generation of dopaminergic neurons from hESCs for therapeutic applications, and a ready source of large numbers of neurons for potential screening applications.
منابع مشابه
Differentiation of human embryonic stem cells into neurons
Human embryonic stem (ES) cells are undifferentiated pluripotent cells derived from the inner cell mass of blastocyst stage embryos. These unique cell lines have the potential to form virtually any cell type in the body and can be propagated in vitro indefinitely in an undifferentiated state. These cells are capable of forming embryoid bodies (EB) that contain cells from all three embryonic lin...
متن کاملDifferentiation of human embryonic stem cells into neurons
Human embryonic stem (ES) cells are undifferentiated pluripotent cells derived from the inner cell mass of blastocyst stage embryos. These unique cell lines have the potential to form virtually any cell type in the body and can be propagated in vitro indefinitely in an undifferentiated state. These cells are capable of forming embryoid bodies (EB) that contain cells from all three embryonic lin...
متن کاملReprogramming by cytosolic extract of human embryonic stem cells improves dopaminergic differentiation potential of human adipose tissue-derived stem cells
The extract of pluripotent stem cells induces dedifferentiation of somatic cells with restricted plasticity. In this study, we used the extract of human embryonic stem cells (hESC) to dedifferentiate adipose tissue-derived stem cells (ADSCs) and examined the impact of this reprogramming event on dopaminergic differentiation of the cells. For this purpose, cytoplasmic extract of ESCs was prepare...
متن کاملEfficient Generation of Functional Dopaminergic Neurons from Human Induced Pluripotent Stem Cells Under Defined Conditions
Human induced pluripotent stem cells (iPSCs) reprogrammed from somatic cells represent a promising unlimited cell source for generating patient-specific cells for biomedical research and personalized medicine. As a first step, critical to clinical applications, we attempted to develop defined culture conditions to expand and differentiate human iPSCs into functional progeny such as dopaminergic...
متن کاملImprovement in Signs of Parkinson's Disease in Rats Following Transplantation of Embryonic Stem Cells
Purpose: Parkinson's disease is a degenerative disease produced by the death of dopaminergic neurons, and the response to current treatments is varied. It is important to develop a model for the evaluation of ES cells as an alternative model for treatment. Materials and Methods: The model for PD was developed in rats. First, ES cells were transplanted into experimental models in three groups: ...
متن کامل