Enhancement of Chondrogenesis of Cultured Quail Limb Bud Mesenchymal Cells by Cellophane Films

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Abstract
Cultures of mesenchymal cells dissociated from the hind limb buds of 3.5 day quail embryos (stages 22-23) were used to study the effect of the cellular microenvironment on chondrogenesis. The degree of chondrogenesis in cultures was estimated by 2 methods. The number of cartilage nodules (characteristic structures formed by chondrocytes) was counted under a phase-contrast microscope, and the amount of toluidine blue absorbed into the extracellular matrix of chondrocytes was measured by spectrophotometry. A culture technique with cellophane films was devised to establish a suitable microenvironment for culture. When mesenchymal cells were cultured by this technique, chondrogenesis was much greater than in usual cultures without cellophane films. Examinations of differences between cultures with and without films indicated that cellophane films promoted differentiation of mesenchymal cells, by the accumulation of some effective factor(s) in culture medium.