Geometric analysis of chondrogenic self-organisation of embryonic limb bud cells in micromass culture.
Item Type:Journal Article
Citation:Rolfe, RA. Shea, CA. and Murphy P., Geometric analysis of chondrogenic self-organisation of embryonic limb bud cells in micromass culture., Cell and Tissue Research, 2022
submitted manuscript Geometric analysis of Micromass Culture.pdf (Accepted for publication (author's copy) - Peer Reviewed) 772.1Kb
Spatial and temporal control of chondrogenesis generates precise, species-specific patterns of skeletal structures in the developing vertebrate limb. The pattern-template is laid down when mesenchymal cells at the core of the early limb-bud condense and undergo chondrogenic differentiation. Although the mechanisms involved in organising such complex patterns are not fully understood, the interplay between BMP and Wnt signaling pathways is fundamental. Primary embryonic limb bud cells grown under high-density micromass culture conditions spontaneously create a simple cartilage nodule pattern, presenting a model to investigate pattern generation. We describe a novel analytical approach to quantify geometric properties and spatial relationships between chondrogenic condensations, utilizing the micromass model. We follow the emergence of pattern in live cultures with nodules forming at regular distances, growing and changing shape over time. Gene expression profiling supports rapid chondrogenesis and transition to hypertrophy, mimicking the process of endochondral ossification within the limb bud. Manipulating the signaling environment through addition of BMP or Wnt ligands, as well as the BMP pathway antagonist Noggin, altered the differentiation profile and nodule pattern. BMP2 addition increased chondrogenesis while WNT3A or Noggin had the opposite effect, but with distinct pattern outcomes. Titrating these pro- and anti-chondrogenic factors and examining the resulting patterns, supports the hypothesis that regularly spaced cartilage nodules formed by primary limb-bud cells in micromass culture is influenced by the balance of Wnt and BMP signaling under a Turing-like mechanism. This study demonstrates an approach for investigating the mechanisms governing chondrogenic spatial organization using simple micromass culture.
Other Titles:Roles of Skeletal Muscle in Organ Development
Type of material:Journal Article
Series/Report no:Cell and Tissue Research
Availability:Full text available