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Application of honeycomb-shaped GelMA microspheres in construction of tumor model

A honeycomb and microsphere technology, applied in the field of biomedicine, can solve the problems of lack of molecular markers in the early diagnosis and prognosis of osteosarcoma, the inability to screen drugs in a repeatable and reliable way, and the increase in tumor drug resistance, etc., to achieve maintenance Biological characteristics and gene stability, saving time cost and economic cost, effect of increasing drug resistance

Pending Publication Date: 2021-12-10
THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The tumor models constructed by three-dimensional culture mainly include tumor cell spheroids and organoids. The former mainly cultures tumor cells on ultra-low cell adhesion plates to form tumor spheroids. However, the 3D tumor models formed by this method often have great differences and cannot be used as reliable Screen Drugs in a Repeatable and Reliable Way
[0006] Osteosarcoma is a common clinical primary malignant bone tumor originating from osteoblasts. At present, there is a lack of ideal molecular markers for the early diagnosis and prognosis of osteosarcoma clinically. no suitable treatment
Simultaneous tumor heterogeneity and increased resistance of recurrent patients to currently used chemotherapeutic drugs are major difficulties in the treatment of osteosarcoma.

Method used

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  • Application of honeycomb-shaped GelMA microspheres in construction of tumor model
  • Application of honeycomb-shaped GelMA microspheres in construction of tumor model
  • Application of honeycomb-shaped GelMA microspheres in construction of tumor model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] The preparation of embodiment 1 honeycomb GelMA microsphere

[0079] Experimental materials: Gelatin was purchased from Rousselot (Zhejiang) Co., Ltd. Methacrylic anhydride was purchased from Aladdin Reagents (Shanghai) Co., Ltd.

[0080] A method for preparing honeycomb-shaped GelMA microspheres for constructing a tumor model, the specific steps are as follows:

[0081] (1) Preparation of GelMA: Dissolve 40.0g of gelatin in 400mL of PBS (50°C), accelerate the dissolution on a magnetic stirrer, add 32.0mL of methacrylic anhydride, and pump it in with a micro-syringe pump while stirring, It is used to mediate the displacement reaction between methacryl groups and amino groups and hydroxyl groups on active amino acid residues on gelatin, and then diluted 5 times with 1600 mL of PBS to terminate the reaction. Dialyze for 2 weeks using a dialysis bag with a molecular weight cut-off of 10 kDa, changing ddH every day 2 O, to completely remove free methacrylic anhydride unt...

Embodiment 2

[0084] Example 2 Exploration of the Characterization of Honeycomb GelMA Microspheres

[0085] The microspheres were observed in general by an inverted fluorescence microscope (bright field) and the particle size distribution of the microspheres was calculated by ImageJ; the freeze-dried microspheres were sprayed with gold by ion sputtering, and then observed under SEM and analyzed by ImageJ Calculate the pore size distribution; observe the surface roughness of the honeycomb GelMA microspheres by AFM.

[0086] 1. Under the scanning electron microscope (SEM), the microstructure of microspheres with GelMA concentration of 5%, 7.5% and 10% were observed respectively, and the diameters were all about 350 μm. However, the microspheres produced by different concentrations of GelMA formed different sizes Pore ​​diameter, the average pore diameter of 5%, 7.5% and 10% microspheres is 25.3±6.5μm, 20.1±5.2μm and 7.9±2.6μm, respectively (supporting information such as figure 1 (A) SEM ima...

Embodiment 3

[0088] Example 3 Further Exploration of the Characterization of Honeycomb GelMA Microspheres

[0089] 1. Use an optical microscope to observe the general view of the microspheres before and after freeze-drying. It can be seen that the microspheres before and after freeze-drying are uniform in size and regular in shape. After contact with water, it will return to the diameter before freeze-drying. This swelling phenomenon provides conditions for the adsorption of cells through swelling during subsequent culture of cells (supporting information such as figure 2 (A), figure 2 (B) shown).

[0090] 2. The microspheres cultured K7M2 cells for 0, 3, and 5 days were dehydrated by ethanol gradient dehydration, and observed under SEM. It can be seen that the microspheres have many pores and are evenly distributed, and the inside and outside are connected, and the cells grow well on the surface of the microspheres And in the pores, the honeycomb-like structure provides more space for...

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Abstract

The invention discloses application of honeycomb-shaped GelMA microspheres in construction of a tumor model. The honeycomb-shaped GelMA microspheres take an isopropyl myristate solution as a continuous phase and a GelMA solution containing 1% of 2-hydroxy-4'-(2-hydroxyethoxy)-2-methyl propiophenone as a dispersed phase, and a honeycomb-shaped structure is formed by freeze-dried GelMA microspheres through preparing monodisperse liquid drops with the same size in batches by adopting a microfluidic technology under a constant speed difference, exposing the liquid drops to 6.9mW / cm<2> ultraviolet light for photo-crosslinking, and carrying out cleaning. According to the invention, honeycomb-like porous GelMA hydrogel microspheres are formed by the microfluidic technology and are applied to three-dimensional culture of osteosarcoma cells. In-vivo and in-vitro experiments show that three-dimensional culture based on the honeycomb-shaped porous microspheres can maintain dryness of the osteosarcoma cells, biological characteristics of the osteosarcoma cells are better maintained, and a real in-vivo environment of the tumor cells can be better simulated.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of honeycomb-shaped GelMA microspheres in constructing tumor models. Background technique [0002] Nowadays, the treatment of malignant tumors includes surgical resection, chemotherapy, radiotherapy and immunotherapy, but the effect of radical cure has not been achieved. For this reason, the development of new treatment modalities is a top priority. The top ten challenges faced by current cancer treatment is how to establish a tumor model that is closer to natural tumors. There are many literatures showing that the treatment methods developed by ourselves have produced excellent therapeutic effects on tumor cell lines and the tumor-bearing animal models modeled by the cell lines, but they have not been applied clinically. The main reason is that tumors Models differ significantly from natural tumors. Therefore, when researchers are looking for w...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C08F299/00C08J9/28C08L55/00
CPCC12N5/0693C12N5/0654G01N33/5011C08F299/00C08J9/28C12N2513/00C12N2533/30C12N2503/02G01N2500/10C08J2355/00
Inventor 施勤崔文国何家辰孙杰
Owner THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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