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Bone accellular stent-based human multiple myeloma micro-environment model building method

A multiple myeloma, decellularized scaffold technology, applied in the direction of cell culture support/coating, animal cells, tumors/cancer cells, etc., can solve the problem of limiting the progression of multiple myeloma, high cost, bone extracellular matrix components unnatural issues

Active Publication Date: 2018-11-13
AFFILIATED HOSPITAL OF NANTONG UNIV
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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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  • Bone accellular stent-based human multiple myeloma micro-environment model building method
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  • Bone accellular stent-based human multiple myeloma micro-environment model building method

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Embodiment Construction

[0033] (1) Preparation of bone decellularized scaffolds

[0034] Take the skull of the healthy adult SD rats that have been killed, remove the soft tissue, and cut the skull into 0.5cm 2 The size of bone slices, the weight is about 15mg / piece. Shake and wash with double distilled water for 1 hour at a speed of 100 rpm. During this period, replace fresh double distilled water several times. After there is no obvious residual blood on the bone slices, wash with 1% SDS + 0.15% ammonia water eluent on a shaker. For 4 weeks, the eluent was changed every 3 days, and the shaker speed was 100 rpm. After 4 weeks, compared with non-decellularized bone slices (see figure 1 ), the acellular bone flakes are relatively transparent (see figure 2 ). Shake and wash with double-distilled water for 1 hour to remove residual eluent on the bone slices. The speed of the shaker is 100 rpm, and fresh double-distilled water is replaced several times during this period. The bone slices were freeze...

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Abstract

The invention discloses a bone accellular stent-based human multiple myeloma micro-environment model building method, which comprises steps of preparation of a bone accellular stent, evaluation of anaccellular effect of the bone accellular stent, cobalt-60 radiation for sterilization, inoculation of human multiple myeloma cells and the like. The invention provides the human multiple myeloma micro-environment model building method to solve the problem that a current multiple myeloma model can hardly simulate a real environment in the body, so that a cell model is provided for further researchon multiple myeloma diseases.

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...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2533/90
Inventor 戚菁李晓红郭益冰徐联成姚俊中陆晶晶申娴娟朱慧鞠少卿顾志峰
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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