Construction method of human multiple myeloma microenvironment model based on bone decellularized scaffold

Abstract

The invention discloses a method for constructing a human multiple myeloma microenvironment model based on a bone decellularized scaffold, including the preparation of the bone decellularized scaffold, the evaluation of the decellularized effect of the bone decellularized scaffold, cobalt 60 Irradiation sterilization, human multiple myeloma cell inoculation and other steps. The invention provides a method for establishing a human multiple myeloma microenvironment model, which solves the problem that the current multiple myeloma model is difficult to simulate the real environment in the body, and provides a cell model for further research on the multiple myeloma disease.

Description

technical field [0001] The invention relates to a method for establishing a human multiple myeloma microenvironment model, and belongs to the technical field of establishing a tumor cell microenvironment model. Background technique [0002] Multiple myeloma (Multiple myeloma, MM) is a malignant B-cell clonal disease with abnormal proliferation of plasma cells, and its tumor cells originate from plasma cells in the bone marrow. tumor. The incidence of this disease has gradually increased in the past few decades, accounting for 10% of all malignant hematological diseases, and the survival rate is only about 35%. . The onset of multiple myeloma is slow, and there are no obvious symptoms in the early stage, so it is easy to be misdiagnosed and missed. [0003] During the development of multiple myeloma, tumor cells interact with their microenvironment. Myeloma cells have a certain effect on the remodeling of the bone environment. Myeloma cells secrete osteoclast activity fac...

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Problems solved by technology

At the same time, because multiple myeloma cells belong to the type of cells that grow in suspension, the growth direction is different from that of adherent cells, showing three-dimensional extension, conventional two-dimensional culture cannot meet its growth characteristics, multiple myeloma cultured in two-dimensional environment The biological properties of cells may be partially changed and cannot reflect the real tumor characteristics in vivo, which will greatly limit the progress of multiple myeloma-related research; the in vivo model of multiple myeloma-an animal model, although close to the tumor in the human body Cell growth status, but the operation is cumbersome, the modeling time is long, the cost is large, and the controllability is poor, which has caused certain difficulties for the downstream mechanism research

At present, people hope to establish a three-dimensional culture model of multiple myeloma in vitro through new technologies to make up for the current deficiencies. In 2018, it was reported that a 3D culture model of multiple myeloma in vitro can be constructed by using bone biomaterials through 3D printing technology, but this method is not suitable for bone biomaterials. The internal microenvironment can only be highly simulated, the extracellular matrix components of bone are not natural, and there is still a certain gap with the real internal bone environment (Braham MVJ, Ahlfeld T, Akkineni AR, MinnemaMC, Dhert WJA, Öner FC, Robin C, Lode A , Gelinsky M, Alblas J. Endosteal and Perivascular Subniches in a 3D Bone Marrow Model for Multiple Myeloma. TissueEng Part C Methods. 2018 Apr 18. doi: 10.1089 / ten.TEC.2017.0467.)

There are also some 3D models of multiple myeloma mainly through the co-cultivation of multiple myeloma cells and one or more other cells in the bone marrow in biomaterials or bioreactors to study the interaction between cells, but Current models cannot be used to study interactions between cells and extracellular matrix components (Belloni D, Heltai S, Ponzoni M, Villa A, Vergani B, Pecciarini L, Marcatti M, Girlanda S, Tonon G, Ciceri F, Caligaris -Cappio F, Ferrarini M, Ferrero E. Modeling multiple myeloma-bonemarrow interactions and response to drugs in a 3D surrogate microenvironment. Haematologica. 2018 Apr;103(4):707-716.)

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