In vitro demonstration of cell-to-cell interaction in growth plate cartilage using chondrocytes established from p53-/- mice.
Three clonal cell lines (MMR14, MMR17, and MMR32) were established from the costal cartilage derived from p53-/- mice. Expression profiles of cartilage-related molecules in MMR14 and MMR17 were compatible with those in cells of the hypertrophic zone. Prolonged in vitro culture induced the expression of calcification-related genes in both cell lines, but calcified nodules were observed only in MMR14. The expression profile of cartilage-related molecules in MMR32 was compatible with that of cells in the perichondrium, with high expression levels of decorin, bone morphogenetic protein-3, and parathyroid hormone-related peptide (PTHrP). When MMR14 was co-cultured with an equal amount of MMR32 without direct contact, the nodule formation was completely inhibited, whereas no such inhibition was observed when MMR14 was co-cultured with MMR17, indicating that soluble factors produced by MMR32 were responsible for the inhibition. Blocking the effects of PTHrP by either antagonizing peptide or neutralizing antibody against PTHrP failed to rescue the inhibitory effects of MMR32, and no increase of the cyclic adenosine monophosphate production in MMR14 was observed when co-cultured with MMR32, suggesting that soluble factors other than PTHrP produced by MMR32 were responsible for the inhibition of terminal differentiation of hypertrophic chondrocytes. This report is the first to show cell-to-cell interaction in the growth plate using cell lines, which will be useful material to investigate the regulatory mechanism of chondrocyte differentiation.
Nakamata
Takeharu
T
Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
Aoyama
Tomoki
T
Okamoto
Takeshi
T
Hosaka
Taisuke
T
Nishijo
Koichi
K
Nakayama
Tomitaka
T
Nakamura
Takashi
T
Toguchida
Junya
J
D000818:Animals; D055398:Bone Morphogenetic Protein 3; D019485:Bone Morphogenetic Proteins / Q000235:genetics; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000502:physiology*; D003094:Collagen / Q000235:genetics; D058575:Decorin; D016326:Extracellular Matrix Proteins; D015870:Gene Expression; D016158:Genes, p53; D006132:Growth Plate / Q000166:cytology* / Q000502:physiology; D051379:Mice; D018345:Mice, Knockout; D044162:Parathyroid Hormone-Related Protein; D010446:Peptide Fragments / Q000494:pharmacology; D036361:Peptide Hormones / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D010641:Phenotype; D011506:Proteins / Q000494:pharmacology; D011509:Proteoglycans / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D055398:Bone Morphogenetic Protein 3; D019485:Bone Morphogenetic Proteins; C546879:Dcn protein, mouse; D058575:Decorin; D016326:Extracellular Matrix Proteins; D044162:Parathyroid Hormone-Related Protein; D010446:Peptide Fragments; D036361:Peptide Hormones; D011506:Proteins; D011509:Proteoglycans; D012333:RNA, Messenger; C054296:parathyroid hormone-related protein (1-34); D003094:Collagen