Isolation, expansion and differentiation of cellular progenitors obtained from dental pulp of agouti (Dasyprocta prymnolopha Wagler, 1831)

Abstract

The study aimed to isolate, expand, differentiate and characterize progenitor cells existent in the dental pulp of agouti. The material was washed with PBS solution and dissociated mechanically with the aid of a scalpel blade on plates containing culture medium D-MEM/F-12, and incubated at 5% CO2-37⁰C. The growth curve, CFU assay, osteogenic/adipogenic differentiation and characterization were obtained from the isolation. The cells began to be released from the explant tissue around the 7th day of culture. By day 22 of culture, cells reached 80% confluence. At the UFC test, 81 colonies were counted with 12 days of cultivation. The growth curves before and after freezing showed a regular growth with intense proliferation and clonogenic potential. The cell differentiation showed formation of osteoblasts and fat in culture, starting at 15 days of culture in a specific medium. Flow cytometry (FACs) was as follows: CD34 (positive), CD14 (negative), CD45 (negative), CD73 (positive), CD79 (negative), CD90 (positive), CD105 (positive), demonstrating high specificity and commitment of isolated cells with mesenchymal stem cells strains. These results suggest the existence of a cell population of stem cells with mesenchymal features from the isolated tissue in the explants of agouti dental pulp, a potential model for study of stem cell strains obtained from the pulp tissue.

Allogeneic mesenchymal stem cells and xenogenic platelet rich plasma, associated or not, in the repair of bone failures in rabbits with secondary osteoporosis

“…According to these authors, xenogeneic PRP has a high immunogenic factor when compared to allogeneic and autogenic presentations. However, being one of the main limitations to the clinical use of PRP, the low yield when compared to the volume of the initial plasma sample 10 , especially for infusion in extensive tissue areas, it becomes urgent to search for alternatives that allow obtaining higher volumes of PRP for continuous use. Due to the scarce studies in human and veterinary medicine on the subject, it was proposed the use of PRP obtained from a large animal in the model of osteoporosis of the present research.…”

“…The cells were isolated from the bone marrow of healthy, expanded in vitro rabbits submitted to osteogenic and chondrogenic cell differentiation assay and aliquoted at the concentration of 1x10 6 cells/mL for immediate use, according to methodologies previously described by Argolo Neto et al 9,16 and Carvalho et al 10 .…”

“…Then, with an orthopedic drill equipped with a 2 mm steel drill and a drill guide 10 , at a speed of 5000 rpm, bone failures of 2 mm in diameter were performed, with two failures in the proximal superior and inferior diaphyseal region of the right tibia and two failures in the proximal superior and inferior diaphyseal region of the left tibia, perpendicular to the bone diaphysis.…”

“…The decalcified fragments were routinely processed, included in histological paraffin 9 , sectioned in a rotating microtome 10 , adjusted to 5μm thickness, stained by Hematoxylin & Eosin and analyzed in binocular optical microscope in x10 and x20 objective.…”

“…been highlighted as an innovative proposal, due to the high plasticity of these cells and the demonstration of in vitro differentiation capacity in osteoblasts 9,10 . Previous studies have already demonstrated the efficacy of MSC in the consolidation of fractures in healthy or non-healthy patients 11,13 .…”