Studies on the transplantation of spinal cord tissue in the rat. I. The development of a culture system for hemisections of embryonic spinal cord

Abstract

This paper describes methods which have been useful in the culture of hemisections of the embryonic rat spinal cord suitable for transplantation into adult rats. The essential feature of the method is the use of small molded fluoroplastic (Aclar) dishes 1 in. in diameter placed in modified petri dishes. These ‘minidishes’ are coated internally with collagen which supports explants grown in a small volume (0.25 ml) of medium in a 5% CO 2 atmosphere. This assembly allows easy access to the tissue, as well as serial high resolution microscopic examination with an inverted compound microscope. Tissue can be fixed, dehydrated and embedded for light or electron microscopy directly in the Aclar minidish. Hemisections of embryonic spinal cord show vigorous outgrowth in this dish and can be maintained for extended periods (weeks) with refeeding twice weekly. Myelinogenesis and synaptogenesis occur in the cord hemisections; when explanted with dorsal root ganglia maturation of the ganglion cells occurs and peripheral myelin is formed. This culture system is also useful for the culture of multiple smaller nervous tissue explants, such as various combinations of spinal cord, cortex or peripheral ganglia.