Growth factors regulate the survival and fate of cells derived from human neurospheres

• Published in: Nature biotechnology Vol. 19; no. 5; pp. 475 - 479
• Main Authors: Caldwell, Maeve A, He, Xiaoling, Wilkie, Neil, Pollack, Scott, Marshall, George, Wafford, Keith A., Svendsen, Clive N.
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.05.2001; Nature; Nature Publishing Group
• Subjects: Aggregates; Agriculture; Animal cells; Animals; Astrocytes - cytology; Astrocytes - drug effects; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Brain - cytology; Brain - embryology; Brain research; Cell Differentiation - drug effects; Cell Division - drug effects; Cell Survival - physiology; Cell Transplantation - methods; Cells, Cultured; Central nervous system; Epidermal growth factor; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Fibroblast Growth Factor 2 - pharmacology; Fundamental and applied biological sciences. Psychology; Growth factors; Growth Substances - pharmacology; Growth Substances - physiology; Humans; Life Sciences; Methods. Procedures. Technologies; Nerve Growth Factors - pharmacology; Neurons - cytology; Neurons - drug effects; neurospheres; Neurotrophic factor 3; Neurotrophic factor 4; Platelet-Derived Growth Factor - pharmacology; Rats; Rodents; Spheres; Stem cells; Stem Cells - cytology; Stem Cells - drug effects; technical-report; Human; Cell culture; Neurotrophic factor; Serum free medium; Cell contact; Neuron; Animal; Nervous system; Cell differentiation; Survival; Platelet derived growth factor
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/88158

Cells isolated from the embryonic, neonatal, and adult rodent central nervous system divide in response to epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF-2), while retaining the ability to differentiate into neurons and glia 1 , 2 . These cultures can be grown in aggregates termed neurospheres, which contain a heterogeneous mix of both multipotent stem cells and more restricted progenitor populations 3 , 4 . Neurospheres can also be generated from the embryonic human brain 5 , 6 , 7 and in some cases have been expanded for extended periods of time in culture 8 , 9 , 10 . However, the mechanisms controlling the number of neurons generated from human neurospheres are poorly understood. Here we show that maintaining cell–cell contact during the differentiation stage, in combination with growth factor administration, can increase the number of neurons generated under serum-free conditions from 8% to >60%. Neurotrophic factors 3 and 4 (NT3, NT4) and platelet-derived growth factor (PDGF) were the most potent, and acted by increasing neuronal survival rather than inducing neuronal phenotype. Following differentiation, the neurons could survive dissociation and either replating or transplantation into the adult rat brain. This experimental system provides a practically limitless supply of enriched, non-genetically transformed neurons. These should be useful for both neuroactive drug screening in vitro and possibly cell therapy for neurodegenerative diseases.