Culture stability of morphogenetic properties of chick limb‐bud mesoderm

Abstract
Stability of morphogenetic properties of leg bud cells (stage 20+) after two culture procedures was tested with a technique described by Zwilling ('64). This technique involves the placing of the cells to be tested into ectodermal jackets obtained from stage 20–21 limb buds and then grafting the composites either to the dorsum of the wing bud or over the somites of a host embryo. There is a decline in the limb properties between 12 and 18 hours of either shaking flask or plate culture. This decline parallels that observed in the limb bud in vivo. The distal area of the wing dorsum as a graft site partially overcomes the decline of the limb properties. Plating at high cell density retards the decline in morphogenetic properties.It is concluded that the culture conditions used in the present experiments have little, if any, effect on the stability of the morphogenetic properties of leg bud cells. Loss of these properties appears to be time dependent. This situation has possible implications for future in vitro studies of the molecular basis for the properties of the morphogenetic phase of chick limb development.