Isolation and Characterization of Female Rabbit Periosteal Derived Progenitor Cells

Abstract

Abstract Background Periosteum is a specialized highly vascularized connective tissue that is formed of an external fibrous layer containing collagen fibers and micro vessels and an inner cambium layer that facing the bone. Cambium layer includes periosteum derived progenitor cells (PDPCs) which have osteochondrogenic potential both in vitro and in vivo and act as major players in bone development and fracture healing. Periosteum derived progenitor cells display properties of mesenchymal stem cells (MSCs), including clonogenicity, phenotypic marker expression, and ability to differentiate into osteoblasts, chondrocytes and adipocytes. So, they are recognized to be essential for the initiation of bone graft healing and remodeling, but their clinical application is still limited due to inaccessibility. The aim of the present study is isolation, culture and characterization of rabbit’s PDPCs. Methods twenty-five adult female rabbits were used to harvest periosteal samples. Cell cultures were obtained by migration of cells from periosteal fragments (explant culture). Immunostaining of the cells of PDPCs in subculture (third passage) was performed. Result On day three of primary culture, the PDPCs were not started to migrate from the periosteal explant edges. However, on day six of primary culture, few spindle PDPCs cells appeared at the edge of explant. on day 20 of primary culture, PDPCs were mostly confluent ( about 9095%) . Immunostaining of PDPCs in subculture (third passage) showed positive brownish reaction of the branched cells for CD105 and CD44 and negative immune reaction for both CD34 and CD45. Conclusion The current study yielded isolation and cultured of female rabbit Periosteal derived progenitor cells. Periosteal derived stem cells are display properties of mesenchymal stem cells (MSCs), including clonogenicity, phenotypic marker expression.