Pluripotent Cell-Derived Glial Precursor Cells for the Delivery of Therapeutic Proteins to the Central Nervous System
Pluripotent stem cells, including human embryonic stem cells and induced pluripotent stem cells, are defined by their ability to differentiate to form any of the cells of the embryo and mature organism, many of which can now be derived in vitro in response to defined factors. Pluripotence, coupled with the ability of these cells to be precisely genetically altered and expanded almost indefinitely, has led to much excitement about their prospects for use in cell transplantation therapies. Cell therapies will require an appropriate source of pluripotent cells that are amenable to expansion to therapeutically useful numbers and robust and reproducible protocols for differentiation of cells to a therapeutically useful phenotype. These protocols will need to involve appropriate quality control measures to ensure the resulting cell populations lack contaminating pluripotent (or other potentially tumorigenic) cells.
In this chapter, we describe the approaches we have used for generating mouse ES cell lines genetically modified to stably express a therapeutic transgene, and the differentiation protocol used to derive highly enriched populations of glial precursor cells from these cells. Discussion is included of some of the measures that have been taken to ensure the functional integrity of these manipulated cell lines is maintained, and the approaches used to determine the purity of the resultant cell populations. Although we have focused on this specific cell type we feel that the structure of this experimental approach could be applied more broadly to deriving other cell types from pluripotent cells.
Development of reproducible protocols for deriving glial precursor cells in large numbers and of high purity will allow further investigation of the specific host dependent and donor cell-dependent factors required for successful cell therapy approaches using various animal models of disease. A detailed understanding of these factors and adaptation of in vitro protocols to human pluripotent cells will ultimately be required for successful clinical translation of cell therapies.
KeywordsPluripotent stem cells Differentiation protocol Glial precursor cell Transgenic protein Stem cell therapy Central nervous system disease
The authors could like to thank Professor John Hopwood, Dr. Kim Hemsley and Professor Peter Rathjen for their support and guidance through the course of these studies. This work would not have been possible without the tireless efforts of Randall and Elizabeth Linton and the generous funding support of the Sanfilippo Children’s Research Foundation.
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