Multiple interactions in juxtaposed monolayers of amphibian neuronal,epidermal, and mesodermal limb blastema cells |
| |
Authors: | Smith Michael J Globus Morton |
| |
Institution: | (1) Department of Biology, University of Waterloo, N2L 3G1 Waterloo, Ontario, Canada |
| |
Abstract: | Summary We have utilized a novel cell culture system to probe nerve-epidermal-blastema cell interactions, in which the cellular participants
are plated as compartmentalized juxtaposed monolayers. A mesoderm monolayer is attached to a culture plate insert and placed
into a tissue culture well, the bottom of which is seeded with a second cell type(s); the two monolayers are separated by
a distance of 1 mm. An examination of the multiple interactions occurring during blastemal growth can thus be accomplished
by deletion of one or more of the cellular participants and employment of a replacement strategy utilizing substances with
putative growth-promoting activity. 3H]Thymidine incorporation into DNA of urodele amphibian blastemal mesoderm cells was evaluated in response to the influence
of juxtaposed brain or epidermal cell monolayers or both cultured in paired combinations. Brain cells were resolved by differential
adhesive selectively into enriched neuronal and glial subpopulations. In the presence of enriched neuronal cells, radiolabel
incorporation into blastemal mesoderm DNA was significantly greater than that achieved by coculturing with glial or unresolved
brain cell populations. The effect of epidermal cells was not overtly expressed in the absence of neuronal cells. The effect
of growth factors fibroblast growth factor (FGF), epidermal growth factor (EGF), nerve growth factor (NGF)] on radiolabel
incorporation into blastemal mesoderm DNA was also assessed in both the presence and absence of neuronal cells. Both EGF and
FGF enhanced radiolabel incorporation into DNA when blastemal mesoderm was cultured in the absence of nerve, but the effect
was significantly less pronounced than that achieved by neuronal cells alone. This system has provided us with an additional
means of characterizing the dialogue between epidermis, nerve, and mesodermal blastema cells, and determining the identity
of growth signals.
This work was supported by National Sciences and Engineering Research Council of Canada grant A6933 to M. Globus. |
| |
Keywords: | regeneration blastema neurotrophic growth factor culture |
本文献已被 SpringerLink 等数据库收录! |
|