Retrovirally transduced bone marrow stromal cells isolated from a mouse model of human osteogenesis imperfecta (oim) persist in bone and retain the ability to form cartilage and bone after extended passaging

Abstract

Bone marrow stromal cells isolated from a model of osteoon the edges of the trabecular bone of the injected and genesis imperfecta (oim) mice, were transduced with a contralateral femurs by X-gal staining and PCR analysis at retrovirus (BAG) carrying the LacZ and neo r genes after 4, 10, 20, 30 and 40 days after injection. The LacZ gene passage 21. The transduced cells retained the ability to was also detected in the lung and liver of the recipient mice express alkaline phosphatase activity in vitro when treated at 4 and 10 days after injection but not at later time-periods. with recombinant human bone morphogenetic protein twoThe present findings suggest that long-term cultured bone (rhBMP-2), formed cartilage in vitro in aggregate cultures marrow stromal cells from osteogenesis imperfecta (OI) and formed bone in ceramic cubes after 6 weeks of implananimals have the potential to traffic through the circulatory tation in nude mice. X-gal staining of ceramic cubes system, home to bone, form bone and continue to express seeded with the transduced cells demonstrated the presexogenous genes. These findings open the possibility of ence of LacZ-positive cells on the edges of bone and also using these cells as vehicles to deliver normal genes to in the lacunae of the newly formed bone 6 weeks after bone as an alternative approach for the treatment of some implantation. After infusion into femurs of oim mice, the forms of OI and certain other bone acquired and genetic transduced cells were detected in the marrow cavity and diseases.

Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

“…These tissues have been demonstrated to contain stem cells of mesenchymal origin, which have potential to differentiate into various specific tissue cell phenotypes that include tendon, fat, marrow stroma, dermis, muscle, bone, and cartilage [10,[12][13][14][15][16][17][18][19][20]. Numerous studies have demonstrated that bone marrow-derived mesenchymal stem cells (MSCs) can give rise to chondrocytes under appropriate conditions [21][22][23]. Members of the TGF-β1 (transforming growth factor 1) super-family have been shown to play a role in the induction of chondrogenic differentiation of MSCs in vitro [21,24,25].…”

Gene therapy approaches for osteogenesis imperfecta

“…38 Several other investigators have reported similar findings in murine, sheep and baboon animal models. [39][40][41][42] Using similar approaches as above but with bettercharacterized mouse bone marrow stromal cells, the current authors have shown that cells infused into mouse femurs persist in bone. Murine adherent marrow stromal cells were established using standard methods and were then marked by transduction with a retrovirus carrying the LacZ marker gene.…”

“…39,43 Infusion of the cells into immunecompetent mice demonstrated that the cells colonized the bones of the recipient mice and differentiated into osteoblasts in vivo. 39,43 Prompted by the murine studies, human clinical trials in children with severe OI were initiated by Horwitz and colleagues using whole marrow harvested from leukocyte antigen (HLA) matched donors. Five children were enrolled in the trial and subjected to irradiation followed by bone marrow transplantation from matched donors.…”

“…Previous studies in mice and humans have demonstrated that bone marrow-derived mesenchymal stem cells will migrate to bone when administered systemically. [37][38][39]46 We previously showed that direct injection of the cells into the mouse bone marrow cavity leads to a wide distribution of the cells in different tissues of the host. 39 Since early treatment for OI will be essential, mesenchymal stem cells transduced with therapeutic genes will need to be delivered early during development.…”

“…[37][38][39]46 We previously showed that direct injection of the cells into the mouse bone marrow cavity leads to a wide distribution of the cells in different tissues of the host. 39 Since early treatment for OI will be essential, mesenchymal stem cells transduced with therapeutic genes will need to be delivered early during development. We have explored this idea by systemic injection of bone marrowderived mesenchymal stem cells transduced with a retrovirus carrying the e-GFP gene (Figure 3a), into the superficial temporal vein of the 2-day-old newborn mice using the methods described previously.…”