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Three-dimensional tissue-like body of composite cells and preparation method and application thereof

A cell, three-dimensional technology, applied in the field of three-dimensional organoids of composite cells and their preparation, can solve the problems of difficulty in nutrient entering into the cell mass, dense cell clumping, and reduced cell survival rate, and achieve the maintenance of cell shape and the improvement of activity. , the equipment requirements and costs are low, and the preparation method is simple and fast.

Inactive Publication Date: 2018-05-29
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Application Information

AI Technical Summary

Problems solved by technology

In the three-dimensional culture without scaffolds, due to the lack of protective support of the scaffold material, the cells may be densely clustered, making it difficult for nutrients to enter the interior of the cell cluster, reducing the cell survival rate

Method used

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  • Three-dimensional tissue-like body of composite cells and preparation method and application thereof
  • Three-dimensional tissue-like body of composite cells and preparation method and application thereof
  • Three-dimensional tissue-like body of composite cells and preparation method and application thereof

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preparation example Construction

[0026] One aspect of the present invention provides a method for preparing three-dimensional organoids of composite cells, the method comprising the following steps: uniformly mixing an aqueous gelatin solution containing a double bond group modification with a suspension of living cells;

[0027] Add photoinitiator and cure by ultraviolet light.

[0028] Preferably, the double bond-containing group-modified gelatin is gelatin modified with acrylic acid or its derivatives. More preferably, the acrylic acid or its derivatives are one or more selected from acrylic acid, methacrylic acid, isopropyl acrylic acid, and methacrylic anhydride. The selection of specific compounds can be carried out according to actual needs. The use of acrylic acid or its derivatives can easily modify the gelatin, thereby cross-linking to build a three-dimensional system. At the same time, the use of acrylic acid or its derivatives to modify the gelatin will not damage the biological activity of the gelatin...

Embodiment 1

[0042] Example 1: Preparation of three-dimensional organoids of compound parathyroid cells

[0043] Among them, the used double bond group-modified gelatin (DB-Gel) and parathyroid cell suspension are prepared as follows:

[0044] A. Preparation of modified gelatin (DB-Gel) containing double bond groups

[0045] Weigh 20g of gelatin and place it in a three-necked flask. Add 200mL of sterile PBS solution and place it on a thermostatic heating magnetic stirrer. Dissolve it at 240r / min and 50℃ for about 2-3h until the pale yellow liquid is clear and uniform without particulate matter. . Connect a constant-pressure burette to the three-necked flask, close the valve, and add 16 mL of methacrylic anhydride; control the dropping rate to one drop every 3-4s, wrap the constant-pressure burette with tin foil, maintain the speed and temperature unchanged, and make the two fully react. About 3-4h. Put an equal volume of PBS solution in a beaker and preheat it to 50°C; after the above reaction...

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Abstract

The invention discloses a three-dimensional tissue-like body of composite cells. By grafting small molecule compounds containing double bonds onto gelatin molecules with excellent biocompatibility tobuild gelatin DB-Gel modified by groups containing double bonds, a gelatin DB-Gel aqueous solution of a certain concentration is mixed with living cell (such as parathyroid gland cells) suspension, and through ultraviolet light curing, the three-dimensional tissue-like body of the composite cells is obtained. In the process of building the tissue-like body, DB-Gel is fully utilized as photo-polymerized hydrogel and has good bioactivity; by directly mixing the gelatin DB-Gel and the cells in a liquid state, the cells can be completely and uniformly dispersed in space, a three-dimensional growing environment closer to the internal environment is built for the cells, the efficiency of interaction between the cells and the external environment is improved in a three-dimensional cell culturingprocess, and compared with cells cultured in two dimensions, maintenance of cell forms and activity are both improved. A preparation method of the three-dimensional tissue-like body of the composite cells is simple, rapid and low in both equipment requirement and cost, and has great potential in internal and external research.

Description

Technical field [0001] The invention relates to the technical field of medical tissue engineering. More specifically, it relates to a three-dimensional organoid of composite cells and a preparation method and application thereof. Background technique [0002] The parathyroid gland is a small endocrine organ in the human body with a simple cell structure, including main cells and eosinophils. Parathyroid cells can secrete parathyroid hormone (Parathyroid Hormone, PTH), which is involved in regulating the balance of calcium and phosphorus in the body. [0003] The interaction between cells and extracellular matrix has a decisive effect on cell fate, so the importance of three-dimensional cell culture is more prominent. The existing explorations on the three-dimensional culture of parathyroid cells mainly include the microencapsulation method and the microsphere carrier method. [0004] The microcapsule embedding method refers to mixing the cell suspension with a biologically active ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N13/00
CPCC12N5/0617C12N13/00C12N2533/54
Inventor 阮长顺李铎梁青飞胡楠张新洲
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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