Comparison of morphological and biochemical characteristics of cultured chondrocytes isolated from proliferative and hypertrophic zones of bovine growth plate cartilage.
Primary cultures of chondrocytes isolated from the proliferative and hypertrophic zones of bovine growth plate cartilage were established, and their morphological and biosynthetic characteristics were compared. High-cell-density monolayer cultures maintained a predominantly chondrocytic morphology, preserved their zone-specific collagen phenotype, and produced an abundant proteoglycan matrix. In contrast, monolayer cultures plated at low cell densities contained many cells with fibroblastic morphology and initiated the synthesis of type I collagen within 72 h following seeding. On a per cell basis, the proliferative zone chondrocytes displayed greater biosynthetic activity than chondrocytes from the hypertrophic region. Characterization of newly synthesized collagens showed that type II collagen was the major biosynthetic product of both types of cells. In contrast, type X collagen was synthesized exclusively by hypertrophic chondrocytes and represented about 20% of the total collagen produced by these cells. The type X collagen molecules synthesized by the hypertrophic cells contained disulfide bonds within their triple-helical domains and assembled into a high-Mr aggregate. When polymerase chain reaction (PCR) was employed to amplify type X collagen sequences from cDNA obtained by reverse transcription of mRNA from cultures of both types of chondrocytes, only the hypertrophic chondrocyte mRNA yielded PCR amplification products indicative of the presence of type X collagen transcripts.
Bashey
R I
RI
Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107.
Iannotti
J P
JP
Rao
V H
VH
Reginato
A M
AM
Jimenez
S A
SA
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003094:Collagen / Q000032:analysis / Q000096:biosynthesis*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006132:Growth Plate / Q000166:cytology / Q000378:metabolism*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000032:analysis
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D003094:Collagen