Chondrogenesis of limb-bud cells: improved culture method and the effect of the potent teratogen retinoic acid

A new apparatus for the mild and reproducible dissociation of rat limb buds into single cells is described. The growth and differentiation of the embryonic mesenchyme cells into chrondrocytes was markedly affected by the conditions of tissue dissociation and the serum used. The degree of chondrogenesis was assessed by alcian blue staining, specific for cartilage matrix. Retinoic acid, a known teratogen, inhibited chondrogenesis completely in a dose-dependent manner. The determination of the quantifiable end-point, IC50 in this cell-culture system, may be suitable to compare the activity of various retinoids and eventually to determine their teratogenic potential.