QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

HOME  |   SEARCH  |   ARCHIVE  |   SUBSCRIBE  |   CONTACT  |   HELP

This Article Full Text (PDF) Submit an eLetter Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by St. John, P. A. Articles by Barker, J. L. Search for Related Content PubMed PubMed Citation Articles by St. John, P. A. Articles by Barker, J. L.

 Previous Article  |  Next Article 

Journal of Neuroscience, Vol 6, 1492-1512, Copyright © 1986 by Society for Neuroscience

ARTICLE

Analysis and isolation of embryonic mammalian neurons by fluorescence- activated cell sorting

PA St. John, WM Kell, JS Mazzetta, GD Lange and JL Barker

Cells were dissociated from the CNS of the embryonic mouse and rat to produce cell suspensions suitable for analysis and separation on a fluorescence-activated cell sorter (FACS). Cells from the spinal cord of the embryonic mouse were analyzed in the most detail. Cell suspensions generated three major peaks in histograms of forward-angle light scatter. Examination of material isolated from each peak and labeling of cell suspensions with the nonvital and supravital fluorescent dyes propidium iodide, ethidium bromide, and acridine orange demonstrated that the three peaks represented live cells, dead cells, and subcellular fragments. Passage through the cell sorter did not detectably damage live cells, as shown by light microscopy, FACS analysis, and in vitro culture of sorted cells. Neurons and glial cells collected by sorting survived at least 4 weeks in culture. Cell suspensions dissociated from the dorsal root ganglia, hippocampus, hypothalamus, cerebellum, and cerebral cortex of the embryonic mouse and from the spinal cord of the embryonic rat produced similar results. Analysis of samples prepared at different developmental stages showed that viable cells could be recovered from each of these regions throughout the important stages of neurogenesis and early cellular differentiation, but that few viable cells could be recovered from animals beyond late embryonic or early postnatal ages. Quantitative FACS analysis of monoclonal antibody A2B5, tetanus toxin and cholera toxin, and lectins binding to live dissociated cells from the embryonic spinal cord demonstrated that these cells had already developed binding sites for these cell-surface ligands by embryonic day 13. These results demonstrate that a fluorescence-activated cell sorter can be used for quantitative analysis of specific cellular properties, that FACS analysis and sorting can be used to identify and isolate live cells from many regions of the embryonic mammalian CNS during important developmental periods, and that sorted neurons and glial cells can be maintained for weeks in culture.

This article has been cited by other articles:

				J. Pruszak, K.-C. Sonntag, M. H. Aung, R. Sanchez-Pernaute, and O. Isacson Markers and Methods for Cell Sorting of Human Embryonic Stem Cell-Derived Neural Cell Populations Stem Cells, September 1, 2007; 25(9): 2257 - 2268. [Abstract] [Full Text] [PDF]

				X. Sun, A. Y. Shih, H. C. Johannssen, H. Erb, P. Li, and T. H. Murphy Two-photon Imaging of Glutathione Levels in Intact Brain Indicates Enhanced Redox Buffering in Developing Neurons and Cells at the Cerebrospinal Fluid and Blood-Brain Interface J. Biol. Chem., June 23, 2006; 281(25): 17420 - 17431. [Abstract] [Full Text] [PDF]

				A.-K. Bouzier, R. Goodwin, F. M.-P. de Gannes, H. Valeins, P. Voisin, P. Canioni, and M. Merle Compartmentation of Lactate and Glucose Metabolism in C6 Glioma Cells. A 13C AND 1H NMR STUDY J. Biol. Chem., October 16, 1998; 273(42): 27162 - 27169. [Abstract] [Full Text] [PDF]

				C.-M. Becker Review : Glycine Receptors: Molecular Heterogeneity and Implications for Disease Neuroscientist, May 1, 1995; 1(3): 130 - 141. [Abstract] [PDF]

Home  |   Search  |   Archive  |   Subscribe  |   Contact  |   Help