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Tissue Preparation and Immunostaining of Mouse Craniofacial Tissues and Undecalcified Bone
Published on: May 10, 2019
Direct comparison of progenitor cells derived from adipose, muscle, and bone marrow from wild-type or
Gregory M Cooper1, Emily L Durham, James J Cray
1Pittsburgh, Pa. From the Departments of Surgery, Oral Biology, Bioengineering, Anthropology, and Orthodontics, University of Pittsburgh; the Pediatric Craniofacial Biology Laboratory, Children's Hospital of Pittsburgh; and the Institute for Complex Engineered Systems, Carnegie Mellon University.
Background:
Reports have identified cells capable of osteogenic differentiation in bone marrow, muscle, and adipose tissues, but there are few direct comparisons of these different cell types. Also, few have investigated the potential connection between a tissue-specific abnormality and cells derived from seemingly unrelated tissues. In this article, the authors compare cells isolated from wild-type rabbits or rabbits with nonsyndromic craniosynostosis, defined as the premature fusion of one or more of the cranial sutures.
Methods:
Cells were derived from bone marrow, adipose, and muscle of 10-day-old wild-type rabbits (n = 17) or from age-matched rabbits with familial nonsyndromic craniosynostosis (n = 18). Cells were stimulated with bone morphogenetic protein-4 (BMP4), and alkaline phosphatase expression and cell proliferation were assessed.
Results:
In wild-type rabbits, cells derived from muscle had more alkaline phosphatase activity than cells derived from either adipose or bone marrow. The cells derived from craniosynostotic rabbit bone marrow and muscle were significantly more osteogenic than those derived from wild-type rabbits. Adipose-derived cells demonstrated no significant differences. Although muscle-derived cells were most osteogenic in wild-type rabbits, bone marrow-derived cells were most osteogenic in craniosynostotic rabbits.
Conclusions:
These results suggest that cells from different tissues have different potentials for differentiation. Furthermore, cells derived from rabbits with craniosynostosis were different from cells from wild-type rabbits. Interestingly, cells derived from the craniosynostotic rabbits were not uniformly more responsive compared with wild-type cells, suggesting that specific tissue-derived cells may react differently in individuals with craniosynostosis.
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