The aim of this investigation was to explore the ex vivo developmental potential of rat limb buds to develop skin in a serum-supplemented organ culture system. Fisher rat fore- and hind-limb buds were microsurgically removed under a dissecting microscope from 13- and 14-day-old embryos and placed on a lens paper supported by a stainless steel grid, where they spent three days or two weeks at the air-liquid interface. Eagle's Minimum Essential Medium was supplemented with 50% rat serum and changed every other day. Samples were processed by routine histology, embedded in paraffin, and uninterrupted serial sections were stained by HE, Masson trichrome or Azan stain. In isolated limb bud, immature epithelium covering its surface was present. During the 3-day culture period, stratified epithelium developed. In limb buds that spent two weeks in culture, keratinization of the stratified epithelium and fully developed stratum granulosum could be discerned in some explants. It is concluded that limb bud organ-culture provides an appropriate model to follow skin development ex vivo, which might be of interest for investigation of pharmacological compounds in skin development.

Temeljne medicinske znanosti