Analysis of chondrogenesis using micromass cultures of limb mesenchyme.
High-density micromass cultures of embryonic mesenchymal cells have proved to be an invaluable model for studying the entire chondrogenic program, from precartilaginous condensations through to chondrocyte hypertrophy. This culture model also provides a powerful system in which to explore the function of various factors in the commitment and differentiation of mesenchymal cells to the chondrogenic lineage. In this regard, micromass cultures provide a consistent and robust model for investigating the effects of genetic manipulations on skeletal phenotypes and for delineating their molecular basis. In this methods chapter, the derivation and use of micromass cultures from murine limb buds are described, but these techniques are also applicable to other organisms and mesenchymal cell sources.
University of British Columbia, Vancouver, BC, Canada.
T Michael
TM
Underhill
University of British Columbia, Vancouver, BC, Canada.
Helen J
HJ
Dranse
University of British Columbia, Vancouver, BC, Canada.
Lisa M
LM
Hoffman
D000818:Animals; D018929:Cell Culture Techniques; D020219:Chondrogenesis / Q000502:physiology*; D005260:Female; D015870:Gene Expression; D006651:Histocytochemistry; D018878:Limb Buds / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D011247:Pregnancy; D014162:Transfection