Sorting of growth plate chondrocytes allows the isolation and characterization of cells of a defined differentiation status.
Axial growth of long bones occurs through a coordinated process of growth plate chondrocyte proliferation and differentiation. This maturation of chondrocytes is reflected in a zonal change in gene expression and cell morphology from resting to proliferative, prehypertrophic, and hypertrophic chondrocytes of the growth plate followed by ossification. A major experimental limitation in understanding growth plate biology and pathophysiology is the lack of a robust technique to isolate cells from the different zones, particularly from small animals. Here, we report on a new strategy for separating distinct chondrocyte populations from mouse growth plates. By transcriptome profiling of microdissected zones of growth plates, we identified novel, zone-specific cell surface markers and used these for flow cytometry and immunomagnetic cell separation to quantify, enrich, and characterize chondrocytes populations with respect to their differentiation status. This approach provides a novel platform to study cartilage development and characterize mouse growth plate chondrocytes to reveal unique cellular phenotypes of the distinct subpopulations within the growth plate.
Belluoccio
Daniele
D
Murdoch Children's Research Institute, University of Melbourne, Royal Children's Hospital, Parkville, Victoria, Australia.
Etich
Julia
J
Rosenbaum
Sabrina
S
Frie
Christian
C
Grskovic
Ivan
I
Stermann
Jacek
J
Ehlen
Harald
H
Vogel
Simon
S
Zaucke
Frank
F
von der Mark
Klaus
K
Bateman
John F
JF
Brachvogel
Bent
B
D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D016923:Cell Death; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D006132:Growth Plate / Q000166:cytology* / Q000254:growth & development; D018189:Immunomagnetic Separation; D051379:Mice; D008810:Mice, Inbred C57BL; D008863:Microspheres; D009928:Organ Specificity; D010641:Phenotype; D015203:Reproducibility of Results
D000954:Antigens, Surface; D015415:Biomarkers