Method for Separating Cells
- *Abstract
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Dr. Steven A. Goldman, M.D., Ph.D., currently Professor and Chairman of the Department of Neurology of the University of Rochester Medical Center, was formerly the Nathan Cummings Professor of Neurology and Neuroscience at Cornell's Weill Medical College. While at Cornell, Dr. Goldman developed several technologies that are available from Cornell for licensing.
Dr. Goldman's laboratory developed a broadly applicable method of isolating prospectively-defined cell types of interest, from both the embryonic and adult nervous systems, using fluorescence-activated cell sorting (FACS). To isolate the desired cell type from a mixed population, a gene encoding a fluorescent reporter protein, such as green fluorescent protein, placed under the control of a promoter or enhancer specifically active in that cell type, is introduced into the cell population. This can be accomplished by viral-mediated transduction (such as by adenoviral infection), electroporation, or liposomal-mediated delivery. Once the targeted cell type, which activates its cognate promoter and hence fluorescent transgene expression, becomes fluorescent, FACS may then be used to separate it from the rest. This method enables the isolation of a wide variety of neural stem and progenitor cells (NSC), as well as their stage-, lineage-, position-, or transmitter-defined progeny, based on the cell-type specific promoters or enhancers employed.
More broadly, this method permits the direct isolation of virtually any cell type for which a gene may be defined that is specifically expressed by that cell type, and for which a cell-specifying promoter or enhancer can be identified. This method has specific advantages in the nervous system, since whereas just a few surface antigens have been identified that are selective for neural stem or progenitor cell types, a large number of cell-type specific genes and their promoters have been identified, that allow a great variety of neural and glial phenotypes and their progenitors to be isolated to purity. Thus, this FACS-based isolation technology is more broadly applicable than antibody isolation.
This method for human neural progenitor cell isolation can thus be used to isolate a variety of lineage-, stage-, transmitter- and position-defined phenotypes, that may be used as targets for screening; as phenotypically-purified preparations for RNA and biochemiocal analyses; as targets for immortalization and subsequent unrestricted expansion; and as cellular vectors for both experimental and therapeutic transplantation. The latter may be particularly appropriate for brain and spinal cord disorders of injury or degeneration in which a single neuronal phenotype (e.g. Parkinson's Disease), glial phenotype (e.g., multiple sclerosis), or small set of discrete cell phenotypes (e.g. segmental spinal cord injury) is affected (see Table 1).
Table 1: Cell Types Capable of Isolation with Specific Promoters
Cell Isolated Promoter(s)(separated by commas) Neurons Enolase, L1 Regenerating neurons MAP-1B Noradrenergic neurons Dopamine beta-hydroxylase Neural or neuronal precursors NCAM Neuronal precursors HES-5 HLH protein, t-alpha-1 tubulin Developing or regenerating neurons alpha-internexin, GAP-43 Peripheral neurons Peripherin Mature neurons Synapsin Oligodendrocytes Cyclic nucleotide phosphorylase I Myelinating oligodendrocytes Myelin basic protein, proteolipid protein Oligodendrocyte precursors Protein JC virus minimal core, cyclic nucleotide phosphorylase II
- *Licensing
- Dan-Oscar Antsonda429@cornell.edu212-746-1297
- 其他
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Goldman SA and Windrem MS, Cell replacement therapy in neurological disease. Phil. Trans. R. Soc. B (2006) 361, 1463-1475
http://www.ncbi.nlm.nih.gov/pubmed/16939969?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum
- 国家/地区
- 美国
