A tissue culture analysis of the steps in limb chondrogenesis |
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Authors: | Michael Solursh Patricia Buckley Ahrens Rebecca S Reiter |
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Institution: | (1) Department of Zoology, University of Iowa, 52242 Iowa City, Iowa |
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Abstract: | Summary Tissue-culture methods can be used to test the developmental capacity of embryonic cells. In micro-mass cultures, derived
from wing cells of stages 21 through 24 chick embryos, aggregates of cells form and then differentiate into cartilage nodules,
as judged by the presence of an Alcian blue staining extracellular matrix. Wing cells derived from embryos as young as stage
17 can form aggregates. However, unless they are treated with db cyclic AMP and theophylline, it is not until stage 20 that
these aggregates can produce cartilage in culture. In clonal cell culture, cartilage colonies are not produced by primary
cell suspensions of limb cells until stage 25 when overt cartilage differentiation is occurring in vivo. It is possible to
obtain clonable cartilage cells from limb cells from embryos between stages 20 and 24 if the cells are either treated with
db cyclic AMP and theophylline or maintained in suspension culture for 12 to 48 hr. On the basis of these in vitro results
a multiple step model for the conversion of limb mesenchyme into cartilage cells is proposed. The model involves the appearance
of cells with a predisposition to form aggregates, development of the capacity to form cartilage in response to elevated levels
of cyclic AMP, the appearance of receptors that translate changes in either cell shape or cell cycle parameters into elevated
levels of cyclic AMP, aggregation, elevated levels of cyclic AMP, cartilage cell determination, and differentiation. This
model can serve as the basis for further tests.
Presented in the Opening Symposium on Nutritional Factors and Differentiation at the 28th Annual Meeting of the Tissue Culture
Association, New Orleans, Louisiana, June 6–9, 1977.
This work was supported by USPHS Training Grant HD00152 from the National Institute of Child Health and Human Development,
while P.B.A. was a postdoctoral trainee, and by NIH Grant HD05505 to M.S. |
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Keywords: | chondrogenesis limb development cell differentiation cyclic AMP |
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