Tissue culture analysis of neurite outgrowth in the presence and absence of serum: possible relevance for central nervous system regeneration.
A tissue culture model has been developed to examine the hypothesis that axons can only regenerate when their growing tips are surrounded by extracellular fluid containing proteins derived from the blood. Fetal rat cerebral explants were cultured in serum medium for 10 days, followed by serum-free (SF) medium (from which serum had been removed) until 18 days in vitro (DIV). All explants cultured in serum medium for 0-10 DIV exhibited greater than 77% neurite viability (neurite viability ratio, NVR, 3.10). This degree of neurite viability was maintained for those explants exposed to serum until 18 DIV (NVR 2.82 at 18 DIV). By contrast, explants maintained in SF medium from 10-18 DIV had a much lower NVR, which, by 18 DIV, had declined to 0.30 (7.5% viability). Transmission electron microscopic analysis of explants fixed at 18 DIV confirmed these phase-contrast results and also showed a predominance of axonal profiles within the neurite population. In the center of explants, tissue viability was in excess of 75% in both the serum and SF media, suggesting that serum is of primary importance for axonal extension rather than neuronal survival. These data strengthen the hypothesis that blood-derived proteins may be needed for prolonged regeneration.
Oorschot
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Jones
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D000818:Animals; D001369:Axons / Q000502:physiology*; D001769:Blood; D002490:Central Nervous System / Q000502:physiology*; D002540:Cerebral Cortex / Q000196:embryology / Q000648:ultrastructure; D046508:Culture Techniques; D009416:Nerve Regeneration; D051381:Rats / Q000196:embryology