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Whole-cell tumor vaccine scaffold and preparation method thereof

A tumor vaccine and whole-cell technology, applied in the field of whole-cell tumor vaccine scaffolds and its preparation, can solve problems such as complicated processes, and achieve the effects of easy acquisition, improved efficiency, and strong antigen immunity

Pending Publication Date: 2022-07-26
WENZHOU INST UNIV OF CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, after the bio-scaffold material is fabricated, additional loading of tumor antigens is often required, which further complicates the process.

Method used

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  • Whole-cell tumor vaccine scaffold and preparation method thereof
  • Whole-cell tumor vaccine scaffold and preparation method thereof
  • Whole-cell tumor vaccine scaffold and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 14

[0020] Example 1 4T1 whole cell tumor vaccine scaffold

[0021] Preparation: as figure 1 As shown, gelatin-methyl methacrylate was dissolved in deionized water at 60°C, completely dissolved and cooled to room temperature. 200 μL of gelatin-methyl methacrylate solution (30% w / v, containing LAP 0.1% w / w), 50 μL of imiquimod solution (240 μg / mL) and 50 μL of mouse breast cancer 4T1 cell solution (4× 10 7 cells / mL) were mixed evenly, poured into a mold, gelatinized after 405 nm light, placed in a -80°C refrigerator for 2 hours, and freeze-dried to obtain a 4T1 whole-cell tumor vaccine scaffold.

[0022] Characterization: The morphology of the 4T1 whole cell tumor vaccine scaffold was observed under the field emission scanning electron microscope, such as figure 2 As shown, macroporous 3D structures and tumor cells are visible in the scaffolds.

[0023] Recruitment and maturation of dendritic cells: 4T1 whole cell tumor vaccine scaffolds and blank scaffolds were implanted into...

Embodiment 2

[0025] Example 2CT26 whole cell tumor vaccine scaffold

[0026] Preparation: Dissolve gelatin-methyl methacrylate in deionized water at 60°C, cool to room temperature after complete dissolution. Formulated Gelatin-methyl methacrylate (10% w / v), hyaluronic acid-methyl methacrylate (10% w / v), HMPP (1% v / v), monophosphoryl lipid A (400 μg / mL), mouse colon cancer CT26 cells (1 × 10 8 cells / mL) mixed solution, poured 300 μL of the mixed solution into a mold, irradiated with ultraviolet light to form a gel, placed in a -20°C refrigerator for 2 hours, and then freeze-dried to obtain a CT26 whole-cell tumor vaccine scaffold.

Embodiment 3

[0027] Example 3 HepG2 whole cell tumor vaccine scaffold

[0028] Chitosan was dissolved in a 0.5% acetic acid solution to obtain a chitosan solution with a concentration of 2wt%, and sodium glycerophosphate was added to adjust the pH value. The gelatin was dissolved in distilled water to obtain a gelatin solution with a solubility of 4 wt%. The gelatin and chitosan were thoroughly mixed 1:1 (v / v). Genipin (0.5w / w), human hepatoma cell HepG2 (1×10 8 cells / mL), CpG oligodeoxynucleotides (250μg / mL) were added to the mixed solution of gelatin and chitosan, 400 μL of the mixed solution was poured into the mold, placed at 37°C to form a gel, and placed in a -80°C refrigerator After 2 hours, the HepG2 whole cell tumor vaccine scaffold was obtained by freeze-drying.

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Abstract

The invention provides a preparation method of a whole-cell tumor vaccine scaffold. The preparation method comprises the following steps: S1, preparing a mixed solution of a scaffold matrix material, a cross-linking agent / photoinitiator, tumor cells and an immunologic adjuvant; and S2, curing the mixed solution obtained in S1, and freeze-drying to obtain the whole-cell tumor vaccine scaffold. The preparation process of the scaffold is simple, and whole-cell tumor vaccine antigen preparation, scaffold preparation and antigen loading are completed through a one-step freeze-drying method. According to the present invention, the freeze-dried tumor cells are adopted as the whole-cell tumor antigen, the immunologic adjuvant is supplemented, and the obtained vaccine scaffold with the macroporous 3D structure can be adopted as the immunogenesis center to recruit the dendritic cells and promote the maturation of the dendritic cells so as to stimulate the anti-tumor immune response, and can be used for preventing the postoperative recurrence and metastasis of the tumor.

Description

technical field [0001] The invention relates to the field of biological materials, in particular to a whole-cell tumor vaccine scaffold based on freeze-drying technology and a preparation method thereof. Background technique [0002] Cancer is a serious threat to human health, and immunotherapy based on tumor vaccines has great development prospects and application potential. Despite the rapid development of tumor vaccines, there are still problems such as limited therapeutic effects and complex production processes. For example, dendritic cell vaccines require cell isolation, modification and screening in vitro, followed by in vivo transplantation, resulting in long production cycles, expensive, and more than 90% of transplanted dendritic cells die before homing to lymph nodes, making Insufficient priming of effector T cells. Tumor cell vaccines, antigen or peptide vaccines, etc. directly activate dendritic cells in vivo, but there are problems such as limited ability to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/54A61L27/56A61L27/36A61K39/00A61K39/39A61P35/00A61P35/04
CPCA61L27/54A61L27/56A61L27/3604A61K39/0011A61K39/39A61P35/00A61P35/04A61L2300/416A61K2039/555A61K2039/55572A61K2039/55561A61K2039/55511A61K2300/00
Inventor 赵远锦邝改真沈贤张庆飞
Owner WENZHOU INST UNIV OF CHINESE ACAD OF SCI
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