Regulation of Axolotl (Ambystoma mexicanum) Limb Blastema Cell Proliferation by Nerves and BMP2 in Organotypic Slice Culture.
We have modified and optimized the technique of organotypic slice culture in order to study the mechanisms regulating growth and pattern formation in regenerating axolotl limb blastemas. Blastema cells maintain many of the behaviors that are characteristic of blastemas in vivo when cultured as slices in vitro, including rates of proliferation that are comparable to what has been reported in vivo. Because the blastema slices can be cultured in basal medium without fetal bovine serum, it was possible to test the response of blastema cells to signaling molecules present in serum, as well as those produced by nerves. We also were able to investigate the response of blastema cells to experimentally regulated changes in BMP signaling. Blastema cells responded to all of these signals by increasing the rate of proliferation and the level of expression of the blastema marker gene, Prrx-1. The organotypic slice culture model provides the opportunity to identify and characterize the spatial and temporal co-regulation of pathways in order to induce and enhance a regenerative response.
Department of Developmental and Cell Biology, University of California Irvine, Irvine, California, United States of America.
Jeffrey
J
Lehrberg
Department of Developmental and Cell Biology, University of California Irvine, Irvine, California, United States of America.
David M
DM
Gardiner
D000558:Ambystoma mexicanum; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D005786:Gene Expression Regulation / Q000187:drug effects; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D018878:Limb Buds / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009417:Nerve Tissue / Q000166:cytology* / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D051785:Smad Proteins / Q000378:metabolism
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D018398:Homeodomain Proteins; C554430:LDN 193189; D011720:Pyrazoles; D011743:Pyrimidines; D051785:Smad Proteins