Organ culture studies on the development of mouse embryo limb buds under EMF influence

Abstract

Purpose: To investigate the effects of an electromagnetic field (EMF) on limb bud development in vitro, an organ culture system was applied.

Materials and methods: Three test groups of amputated mouse limb buds included the experimental (E) group which received EMF (50 Hz/13.1 mT, for 2 h), a sham (Sh) group exposed to no EMF treatment and the control (C) group. The limb buds of E and Sh groups (n = 20 per group) were amputated from mouse embryos on day 11.5 of development and cultured in minimum essential medium Eagle (MEM Eagle), supplemented with 15% human embryo cord serum, for 2 days, while those of group C (n = 20) were removed on day 13.5 of development. All samples were fixed in Bouin's fluid, embedded in paraffin, serially sectioned (5 μm thick) and stained with Hematoxylin and Eosin. Limb bud measurements were performed using a scaled graticule.

Results: Morphological and histological examinations showed significant changes in the experimental limb bud group as compared with the sham and control groups. The growth rate in both fore and hindlimb buds in proximal-distal (P-D) and anterior-posterior (A-P) axes were significantly increased. Chondrocyte counts and mitotic figures of mesenchymal and red blood cells were significantly increased as compared with those of sham and control groups. There was also a significant reduction of mesenchymal cell counts, while no significant difference was observed in the degenerated cell counts among the three groups.

Conclusions: These findings suggest that EMF, under the conditions applied, has progressive effects on the limb bud development and that both proliferation and differentiation can be stimulated in vitro.

Keywords:

• Mouse limb bud
• organ culture
• EMF