Project description:Proliferative zone chondrocytes were microdissected from control and Ift88-deleted growth plates to determine gene expression profiles regulated by primary cilia.
Project description:Proliferative zone chondrocytes were microdissected from control and Ift88-deleted growth plates to determine gene expression profiles regulated by primary cilia. Four total samples were analyzed. Two biological replicates of proliferative zone chondrocytes microdissected from control mice and two biological replicates from Ift88 deleted (Col2aCre;Ift88fl/fl) mice. Control and experimental mice were in the Bl/6 background.
Project description:To study the role of Nox2 in ethanol toxicity on the bone growth plate chondrocytes, we generated Nox2 conditional knockout mice (CKO), in which the catalytic subunit of Nox2, Cybb, is deleted in chondrocytes using a Cre-lox system, where Cre is expressed from the Col2a1 promoter. CKO mice and floxed control mice were fed an ethanol-containing Lieber De-Carli-based diet or pair-fed a control diet for 8 weeks starting at 5-6 weeks of age. As both the Nox2 genotype and ethanol diminished the number of chondrocytes in the growth plates, we conducted an RNA-Seq analysis of the growth-plate containing regions of the femurs.
Project description:Longitudinal bone growth depends upon the execution of an intricate series of cellular activities by epiphyseal growth plate chondrocytes. In order to better understand these coordinated events, microarray analysis was used to compare gene expression in chondrocytes isolated from the proliferative and hypertrophic zones of the avian growth plate. In this experiment we compared pooled samples of proliferative and hypertrophic chondrocytes isolated from the chick growth plate. The expression of 745 genes was found to differ 3-fold or greater at the 0.05 level of probability.
Project description: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.