Resumen
Method to enhance the osteogenic differentiation of mesenchymal stromal cells (MSC). The invention relates to a method for enhancing the osteogenic differentiation of MSCs by culturing them in osteogenic medium with the addition of an EphB3 receptor blocking reagent. Said blocking reagent can be selected from polyclonal antibodies, monoclonal antibodies, EphB3 blocking peptides, fusion proteins, RNAi and/or shRNA blocking the expression of the EphB3 gene, editing of the gene encoding EphB3 using the CRISPR/CAS technique or combinations of the same. The invention also relates both to the cells of the osteogenic lineage obtained by the method described and to the EphB3 receptor blocking reagents, for their use in the treatment of bone pathologies with bone tissue deficiency and/or bone fractures. Pharmaceutical compositions containing the cells of the osteogenic lineage obtained by the described method and/or the EphB3 receptor blocking reagents are also included. (Machine-translation by Google Translate, not legally binding)
Reivindicaciones
1. A method for enhancing the osteogenic differentiation of mesenchymal stromal cells (MSCs) isolated and cultivated in vitro and obtaining cells of the osteogenic lineage, which includes the following steps: 1) after reaching 80-90% confluence between cells, replacing the culture medium used for culturing MSCs with an osteogenic medium and adding an EphB3-receptor blocking reagent; 2) incubating for at least 7 days. 2. The method for enhancing the osteogenic differentiation of MSCs according to claim 1, wherein the in vitro culture of isolated MSCs is performed in a culture medium which favors MSC proliferation. 3. The method for enhancing the osteogenic differentiation of MSCs according to any of the preceding claims, wherein step 1) is performed when the cells reach 90% confluence. 4. The method for enhancing the osteogenic differentiation of MSCs according to any of the preceding claims, wherein the osteogenic medium with the EphB3-receptor blocking reagent is renewed at least every 7 days. 5. The method for enhancing the osteogenic differentiation of MSCs according to any of the preceding claims, wherein the cells are from a mammal. 6. The method for enhancing the osteogenic differentiation of MSCs according to claim 5, wherein the mammal is a human. 7. MSC cells obtained according to the method defined in any of claims 1-6 for use in the treatment of bone diseases with bony tissue deficiency and/or bone fractures. 8. MSC cells obtained according to the method defined in any of claims 1-6 for use according to claim 7, wherein the bone diseases with bony tissue deficiency belong to the group comprising: primary osteoporosis, drug-induced osteoporosis, osteopenia, osteogenesis imperfecta, chronic kidney disease, hyperthyroidism, celiac disease. 9. An EphB3-receptor blocking reagent, element, compound, molecule, and/or modified molecule for use in the treatment of bone diseases with bony tissue deficiency and/or bone fractures. 10. The EphB3-receptor blocking reagent, compound, and/or molecule according to claim 9, selected from the group consisting of: polyclonal antibodies, monoclonal antibodies, fusion proteins, or combinations thereof. 11. The EphB3-receptor blocking reagent, compound, and/or molecule according to any of claims 9-10, wherein the bone diseases with bony tissue deficiency belong to the group comprising: primary osteoporosis, drug-induced osteoporosis, osteopenia, osteogenesis imperfecta, chronic kidney disease, hyperthyroidism, celiac disease. 12. The EphB3-receptor blocking reagent, compound, and/or molecule according to any of claims 9-11 for use in a mammal. 13. The EphB3-receptor blocking reagent, compound, and/or molecule according to claim 12, wherein the mammal is a human. 14. A pharmaceutical composition including an EphB3-receptor blocking reagent, element, compound, molecule, and/or modified molecule for use in the treatment of bone diseases with bony tissue deficiency and/or bone fractures. 15. The pharmaceutical composition according to claim 14, wherein the EphB3-receptor blocking reagent, element, compound, molecule, and/or modified molecule is selected from the group consisting of: polyclonal antibodies, monoclonal antibodies, fusion proteins, or combinations thereof. 16. A pharmaceutical composition including cells of the osteogenic lineage obtained according to the method defined in any of claims 1-6. 17. The pharmaceutical composition according to any of claims 14-15 including cells of the osteogenic lineage obtained according to the method defined in any of claims 1-6. 18. The pharmaceutical composition according to any of claims 14-17, wherein the bone diseases with bony tissue deficiency belong to the group comprising: primary osteoporosis, drug-induced osteoporosis, osteopenia, osteogenesis imperfecta, chronic kidney disease, hyperthyroidism, celiac disease.
