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. 2022 Mar 5;12(5):e4339.
doi: 10.21769/BioProtoc.4339.

Skeletal Stem Cell Isolation from Cranial Suture Mesenchyme and Maintenance of Stemness in Culture

Takamitsu Maruyama  1   2   3 , Hsiao-Man Ivy Yu  1   3 , Wei Hsu  1   4   3   5   6   7
Affiliations

Skeletal Stem Cell Isolation from Cranial Suture Mesenchyme and Maintenance of Stemness in Culture

Takamitsu Maruyama et al. Bio Protoc. .

Abstract

Skeletal stem cells residing in the suture mesenchyme are responsible for calvarial development, homeostatic maintenance, and injury-induced repair. These naïve cells exhibit long-term self-renewal, clonal expansion, and multipotency. They possess osteogenic abilities to regenerate bones in a cell-autonomous manner and can directly replace the damaged skeleton. Therefore, the establishment of reliable isolation and culturing methods for skeletal stem cells capable of preserving their stemness promises to further explore their use in cell-based therapy. Our research team is the first to isolate and purify skeletal stem cells from the calvarial suture and demonstrate their potent ability to generate bone at a single-cell level. Here, we describe detailed protocols for suture stem cell (SuSC) isolation and stemness maintenance in culture. These methods are extremely valuable for advancing our knowledge base of skeletal stem cells in craniofacial development, congenital deformity, and tissue repair and regeneration.

Keywords: Bone regeneration; Calvaria; Cell-based therapy; Craniofacial; Mesenchymal stem cell; Osteogenesis; Skeletal stem cell; Skeletogenic mesenchyme; Sphere culture; Suture stem cell.

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Conflict of interest statement

Competing interestsThe authors declare no competing financial interests.

Figures

Figure 1.
Figure 1.. Schematic of mouse calvarial bones and sutures.
F, frontal bone; P, parietal bone; IP, interparietal bone.
Figure 2.
Figure 2.. Schematic diagrams illustrating the separation of bone pieces to release the cells from the suture mesenchyme.
Figure 3.
Figure 3.. Representative images showing cells after 1 day (A) and 8 days (B) of the sphere culture.
Scale bars, 200 µm.
See this image and copyright information in PMC

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