Establishment Of Human Multiple Myeloma Model And Investigation Of Its Special Quality

Multiple myeloma is a malignant plasma cell disorder. And it is characterized by heterogeneous anemia, bone disease, renal impairment, hypercalcaemia and infections due to patchy accumulation of malignant plasma cells within the marrow microenvironment. With its rather lower incidence in our country MM is still deleterious for patients as its elusive mechanism and incurability. To establish a perfect MM model is so significant as to further research and seek after MM's pathogenesis and investigate new drugs against it.Bone transplantation has often been used for repairing skeletal defect, restoring structural integrity or reconstruction the joint of osteoarthrosis making it broad application in musculoskeletal surgery, oral and maxillofacial surgery, brain surgery, cosmetic and plastic surgery. It contains practicably autologous and allogeneic bone transplantation in addition to heterogeneous bone transplantation applied occasionally. But it is very significant to establish chimera of human and nude mice by utility of the osseous xenograft techniques for investigation of hominine physiological and pathological process of blood system.The presently available mouse models of human PCN can be divided into de novo and transplantation models. Although none of the genetically engineered strains recapitulate all features of a particular human PCN, several strains have emerged as useful platforms for mechanistic and therapeutic studies of alterations in signaling pathway found in human PCN (e.g., IL-6, Abl, and Myc). Nonetheless, they are highly valuable for many other purposes including preclinical drug testing.Recently Yaccoby and his colleagues have developed a classic chimeric SCID-human MM model which could greatly simulate almost all kinds of pathological manifestation and clinic symptoms during the course of MM. Classical SCID-hu model virtually made it as a chimeric niche of human and SCID mice. This kind of niche was characterized of miscellaneous trait at all. So the survival of grafted bone was inevitably associated with various processes in the chimeric niche. Bypast animal models were based on the SCID mice which are hard to bring immunoreaction but they had neglected a fact that MM strikes men with immunity, so it is deficient for them to establish MM model in SCID mice without immunity.Under ground of osseous xenograft techniques, by means of similar approach to the way of building SCID-hu model, also based on the novel BALB/c-nu-hu chimeric niche, we successfully develop a model of human multiple myeloma and investigate its special quality further.Objective: To establish a novel model of human multiple myeloma (MM) and investigate its special quality compared with classical models.Methods: Subcutaneously transplantation of 1cm long segment of 4 weeks old human fetal thigh or tibia bone into BALB/c mice; Make apoptosis check of all cells collected from fetal bone marrow tissue marked by rh Annexin V/PI before and after transplantation both by flow cytometry and fluorescence microscope; Launch Wright-Giemsa dying and paraffin embedding and secting followed by hematoxylin and eosin staining of fetal bone marrow tissue before and after transplantation for morphological examination. Develope a novel model of human multiple myeloma based on the BALB/c-nu-hu chimera and then inoculate the bone marrow mononuclear cells (BMNCs) harvested from primary MM patients into the FBs grafted into the BALB/c nude mice. The level of human immunoglobulin G detected by the Human Ig enzyme-linked immunosorbent assay (ELISA) kit. Besides FBs implanted, important organs of the model mice respectively stained by hematoxylin and eosin (H&E) and monoclonal mouse anti-human CD34 CD59 and CD138 followed by morphological examination. FBs and murine stem bone stained by tartrate-resistant acid phosphatase (TRAP) and examined too. Take X-ray film of model mice at start and terminal of the experiment.Results: Fetal bone can survive in the subcutaneous site of BALB/c nude mice and bear high livability; Quantity and morphology of fetal bone marrow tissue and cells before and after transplantation were close to each other. The multiple myeloma cells (MCs) of primary MM patients can grow, infiltrate and migrate. The model charactered by cachexia, idiosyncratic rather highly level of human IgG, increased osteoclast activity and resorption of the human bones, terminal scatter and migration of human MCs.Conclusion: The novel BALB/c-nu-hu chimeric niche is a sort of credible vector for researching hominine hematopoiesis and bone based on animal. The MM model can be built successfully based on the BALB/c-nu-hu chimera. By comparison with classical ones reported before our model have many virtues like similar efficiency, rather easygoing necessity, high repetition, probably more approaching hominine realistic imitation.