key: cord-0041897-tjwoh16k
authors: ARMSTRONG, R.; FRIEDRICH, V. L.; HOLMES, K. V.; DUBOIS‐DALCQ, M.
title: Proliferation and Differentiation of Neuroglial Cells Isolated during Demyelination and Remyelination
date: 2006-12-17
journal: Ann N Y Acad Sci
DOI: 10.1111/j.1749-6632.1990.tb42429.x
sha: 48b0518d3704ac17ba44bc2e34b2ae1284899b32
doc_id: 41897
cord_uid: tjwoh16k

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We have studied the growth and differentiation properties of oligodendrocytetype 2 astrocyte (0-2A) lineage cells isolated from mouse spinal cord in the course of demyelination and remyelination. When infected at 28 days of age with a coronavirus (murine hepatitis virus strain A59), C57BI/6N mice develop numerous foci of demyelination in the spinal cord within 3-5 weeks postinfection (wpi); extensive remyelination occurs in the following weeks.'

Neuroglial cells were isolated from spinal cords by sequential enzymatic and mechanical dissociation. Glial cells were separated from the myelin and red blood cells by centrifugation through a Percoll gradient. Using three-color immunofluorescence we could simultaneously identify oligodendrocytes with antigalactocerebroside, astrocytes with antiglial fibrillary acidic protein, and all 0-2A lineage cells, including 0-2A adult progenitors,* with the 0 4 monoclonal antibody.

Control cultures contained scattered 0-2A lineage cells, clusters of type 1 astrocytes, and occasional fibroblasts at 1 day in uitro. The 0-2A lineage population was comprised mainly of oligodendrocytes (04+ GC+ GFAP-), although 0-2A adult progenitor cells (04+ GC-GFAP-) and type 2 astrocytes ( 0 4 + GC-GFAP+) were also present. Compared to cultures of control tissue, cultures of demyelinated spinal cord (3-5 wpi) exhibited a striking increase in the total number of 0-2A lineage cells. Additionally, phagocytic cells with characteristics of microglia and/or macrophages were prevalent in these cultures. The 0-2A lineage population cultured from demyelinated tissue consisted of a higher proportion of type 2 astrocytes and cells of a mixed oligodendrocyte-astrocyte phenotype (04+ GC+ GFAP+). 0-2A lineage cells with this mixed phenotype do not appear to be an artifact of the tissue culture conditions, because similar cells have been described in ~i t r o .~

The antigenic phenotypes of 0-2A lineage cells from demyelinated tissue could be influenced in vitro by adding growth factors to the culture medium from 1-3 days in uitro (FIG. 1) . Addition of insulin-like growth factor 1 (IGF-1) increased the relative abundance of oligodendrocytes, whereas basic fibroblast growth factor (FGF) seemed to inhibit galactocerebroside expression. Exogenous platelet-derived growth factor (PDGF) did not modify the relative proportion of oligodendrocytes in these cultures.

We determined the mitogenic potential of 0-2A lineage cells by combining three-color immunofluorescence with tritiated thymidine autoradiography . Tritiated thymidine was administered as either a 2-hour in uiuo pulse or a 20-hour in uitro pulse. Following either labeling protocol, cultures derived from demyelinating tissue ( In this in uirro system, we can now characterize the factors responsible for the phenotypic plasticity and proliferation of 0-2A lineage cells which may underlie the successful remyelination observed in this experimental model of virally induced demyelination.

Expression of viral and myelin gene transcripts in a murine demyelinating disease caused by a coronavirus

Identification of an adult-specific glial progenitor cell

In viuo analysis of glial cell phenotypes during a viral demyelinating disease in mice