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High glucose concentration inhibits migration of rat cranial neural crest cells in vitro

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Summary

Cranial neural crest cells give rise to a large part of the facial structures, and disturbed development of these cells may therefore cause congenital malformations affecting the head and face. We studied the effects of increased glucose concentration on the migration and development of cranial neural crest cells, maintained in vitro for 48 h. Pre-migratory cranial neural crest cells were removed from embryos of normal and diabetic rats on gestational day 9. After 24 h in 10 mmol/l glucose the cells were exposed to glucose concentrations of 10, 30, or 50 mmol/l for another 24 h. The cultures were photographed at 24 h and 48 h in a phase-contrast microscope to evaluate cell morphology, cell number, and cell migration. Exposure to 50 mmol/l glucose reduced the total number of neural crest cells, their mean migratory distance and migratory area expansion compared to cells cultured in 10 mmol/l glucose. To investigate the effect of antioxidant agents, high glucose cultures were studied after addition of N-acetylcysteine (NAC), or Superoxide dismutase (SOD). Addition of NAC diminished the inhibitory effect of high glucose, whereas SOD did not offer any improvement in cell development. Neural crest cell culture from embryos of diabetic rats showed reduced cell migration in vitro at all glucose concentrations compared to normal cells. In addition, the cells from embryos of diabetic rats showed reduced migratory area expansion after culture in the basal 10 mmol/l glucose concentration, indicating that maternal diabetes permanently influences the future development of premigratory cranial neural crest cells. These findings indicate that high glucose concentration inhibits cranial neural crest development in vitro, and that antioxidant therapy may diminish this inhibition. Free radical oxygen species may be involved in the induction of malformations and antioxidants may therefore have a role in future attempts to block the teratogenic effects of diabetic pregnancy.

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Abbreviations

NAC:

N-acetylcysteine

SOD:

Superoxide dismutase

DIC:

difference interference contrast

MCN:

migratory cell number

δ MCN:

increase of MCN during the second 24-h culture period

MMD:

mean maximal distance from the centre of the explant to the edge of the area covered by the migrating cells

δ MMD:

increase in MMD during the second 24-h culture

MTA:

total area covered by the migrating cells

MTA%:

MTA after 48-h of culture in percent of MTA at 24-h of culture

ANOVA:

analysis of variance

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Authors and Affiliations

  1. Department of Medical Cell Biology, University of Uppsala, Uppsala, Sweden

    N. Suzuki & U. J. Eriksson

  2. Department of Developmental Biology and Genetics, University of Uppsala, Uppsala, Sweden

    K. Svensson

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  1. N. Suzuki
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  2. K. Svensson
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  3. U. J. Eriksson
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Suzuki, N., Svensson, K. & Eriksson, U.J. High glucose concentration inhibits migration of rat cranial neural crest cells in vitro. Diabetologia 39, 401–411 (1996). https://doi.org/10.1007/BF00400671

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  • DOI: https://doi.org/10.1007/BF00400671

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