Project Details:
DEVELOPMENT OF HUMAN EMBRYONIC NEURAL PRECURSOR STEM CELLS: BIOLOGY, PHYSIOLOGY, AND TOXICOLOGY
The way in which the various elements of the nervous system develop, integrate, and operate is fundamental to our understanding of how the nervous system functions in both in health and disease. A variety of studies have shown that the developing nervous system can be readily altered, often in quite subtle ways, but that these alterations can affect both the normal adult functioning of the nervous system, and may even play a role in the development of disorders of ageing such as Parkinson’s disease and Alzheimer’s disease. Our aim is to use stem cell methods to understand how neurones from different regions of the human brain function, and also how they become dysfunctional and degenerate. This will provide us with an increased understanding of how the nervous system functions and may also provide us with clues to the aetiology of a wide variety of neurological disorders.
To allow us to study human neural development, differentiation and cell death we have derived human Neural Precursor Cell (hNPC-“stem cell”) lines and are utilising these to study how human neurones develop and function, and also how they are affected by a variety of treatments and manipulations. These hNPC lines can be maintained in serum free defined media as “neurosphere” cultures for periods in excess of a year and can be successfully regrown from liquid nitrogen storage providing a long term supply. Cultures can be induced to differentiate into neurones, astrocytes, and oligodendrocytes by simple growth factor withdrawal and serum addition, or can be directed to differentiate into specific cell types (e.g. dopaminergic neurones) by addition of specific differentiation factors. Once differentiated, these cell lines are stable and show minimal cell division over several weeks, express neuronal (e.g. Gad67) and glial (e.g. GFAP) antigens, and are killed by a range of known neurotoxins (e.g. MPP+). Our studies on hNPCs may therefore provide suitable cell systems to further our understanding of basic human neural development in both health and disease.
Affiliation: Medical Toxicology Centre (http://www.ncl.ac.uk/mtc/)
Institute of Neuroscience (http://www.ncl.ac.uk/ion/)
Institute of Cellular Medicine (http://www.ncl.ac.uk/icm/)
Institute for Ageing and Health (http://www.ncl.ac.uk/iah/)
Research StaffDr Chris Morris
Prof PG Blain
Immunocytochemical analysis of differentiated Human Neural Precusor (Stem) Cells. Cells were grown on culture slides and stained for neurone specific beta-Tubulin (Tuj1 (A)), tyrosine hydroxylase (TH) as a marker of dopaminergic neurones (B), glial fibrillary acidic protein (GFAP) to demonstrate astrocytes (C), and Nestin to show intermediate filament proteins indicating stem cells (D) following exposure to medium containing differentiating agents during a 7 day differentiation period. (×10 magnification).
