Abstract
As a basic science, craniofacial research embraces multiple facets spanning from molecular regulation of craniofacial development, cell biology/signaling and ultimately translational craniofacial biology. Calvarial sutures coordinate development of the skull, and the premature fusion of one or more, leads to craniosynostosis. Animal models provide significant contributions toward craniofacial biology and clinical/surgical treatments of patients with craniofacial disorders. Studies employing mouse models are costly and time consuming for housing/breeding. Herein, we present the establishment of a calvarial suture explant 2-D culture method that has been proven to be a reliable system showing fidelity with the in vivo harvesting procedure to isolate high yields of skeletal stem/progenitor cells from small number of mice. Moreover, this method allows the opportunity to phenocopying models of craniosynostosis and in vitro tamoxifen-induction of ActincreERT2;R26Rainbow suture explants to trace clonal expansion. This versatile method tackles needs of large number of mice to perform calvarial suture research.
Highlights
Calvarial sutures are soft fibrous tissue joining the five flat intramembranous bones of the skull vault (Opperman, 2000; Lenton et al, 2005; Rice, 2008)
In the first part of this manuscript, we will describe the preparation of calvarial sutures explanted from postnatal day 3 and 15 mice, their 2-D in vitro growth and the isolation of a skeletal stem/progenitor cells previously isolated from in vivo calvarial sutures (Menon et al, 2021)
Having established the ex vivo calvarial suture 2-D culture system, we investigated whether this method could be employed to isolate higher or similar yields of skeletal stem/progenitor cells while scaling down the number of mice needed for the in vivo isolation procedure
Summary
Calvarial sutures are soft fibrous tissue joining the five flat intramembranous bones of the skull vault (Opperman, 2000; Lenton et al, 2005; Rice, 2008). These fibrous joints consisting of non-ossified mesenchymal cells (suture mesenchyme) are located between the two approaching osteogenic fronts of the skull vault bone. They serve as growth centers playing critical roles in facilitating the expansion and development of the postnatal skull vault in order to accommodate the growing brain (Morriss-Kay and Wilkie, 2005; Nieman et al, 2012).
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