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A kind of 3D cell culture material and its application

A cell and culture medium technology, applied to the material of three-dimensional culture of cells in vitro and its application field, to achieve the effect of facilitating cell degumming and rapid growth

Active Publication Date: 2018-01-23
GUANGZHOU SAGENE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is still no cell scaffold that can fully meet the rapid and high-viability growth of various cells at the same time, and can meet the needs of stem cells to effectively maintain cell stemness during the proliferation process

Method used

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  • A kind of 3D cell culture material and its application
  • A kind of 3D cell culture material and its application

Examples

Experimental program
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Effect test

preparation example Construction

[0045] PEG8000@Fe 3 o 4 The preparation method:

[0046] 1) Alkylation: take Fe with a particle size of 15-25nm 3 o 4 Nanospheres, add ammonia solution of organic matter, mix well, and react at 8°C-10°C for 16-20h to make Fe 3 o 4 The surface of nanospheres is alkylated by cations to form a protective film;

[0047] 2) Alkylated Fe 3 o 4 Mix nanospheres with phosphorylated PEG8000 aqueous solution, and remove oxygen through argon, then add metal salt, mix well and stir at 55-65°C for 1-2 hours to obtain PEG8000@Fe 3 o 4 ;

[0048] PEG400@Fe 3 o 4 The preparation method of the above-mentioned PEG8000@Fe 3 o 4 The preparation method is the same, except that the phosphorylated PEG8000 is changed to phosphorylated PEG400, and the others are unchanged.

[0049] Preferably, the above-mentioned three-dimensional cell culture material also includes nanoparticles NH 2 @Fe 3 o 4 , its preparation method is:

[0050] 1) Alkylation: take Fe with a particle size of 100-20...

Embodiment 1

[0071] Embodiment 1 A kind of preparation method of three-dimensional cell culture material

[0072] 1) Take Fe with particle sizes of 20nm and 100nm respectively 3 o 4 nanospheres, and Fe 3 o 4 The saturation magnetization of nanospheres is 71emu / g and above; 3 o 4 The nanospheres were added to the ammonia solution with a concentration of 200g / LOleic acid (cis-octadec-9-enoic acid), and at 8°C to 10°C, the reductive catalytic reaction was maintained for 16 to 20h by continuously blowing argon, so that the nanospheres The surface of the ball is alkylated by cations to form a protective film, and the ammonia water is washed away after the reaction is complete;

[0073] 2) Get 100g of alkylated Fe with a particle size of 20nm 3 o 4 Add 400mL (so that the content of the ball is 25%) PEG8000 phosphorylated PEG8000 aqueous solution with a concentration of 0.25g / mL to the nanospheres (that is, PEG8000 and phosphoric acid are added into the water at a molar ratio of 1:1, and t...

Embodiment 2

[0080] Example 2 Application of a three-dimensional cell culture material

[0081] In this example, the 3D cell culture material prepared in Example 1 above is used to culture mesenchymal stem cells (MSC), and the specific steps are as follows:

[0082] 1) Preparation of cytokine nanoparticles: take 0.025g of the nanoparticles NH prepared in Example 1 2 @Fe 3 o 4 Add to 1mL of 4% w / v formaldehyde solution, mix well, then add 2mg cytokine LIF, mix well, and react for 2 hours to get cytokine nanoparticle LIF@Fe 3 o 4 . (According to the specific cells cultivated, it can be prepared into other cytokine nanoparticles; while some cells are cultured without cytokines, the preparation of this step of cytokine nanoparticles can be omitted. In this embodiment, it is to expand the mesenchymal The proliferation and culture of stem cells MSC, cytokine LIF can inhibit the differentiation of stem cells MSC, so the preparation of nanoparticles LIF@Fe 3 o 4 It can effectively maintain ...

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Abstract

The invention discloses a 3D (three-dimensional) cell culture material and application thereof. The 3D cell culture material comprises PEG (polyethylene glycol) 8000@Fe<3>O<4>, PEG400@Fe<3>O<4> and NH2@Fe<3>O<4>. The 3D cell culture material and the application have the advantages that the pH (potential of hydrogen) of micro-environments and the activity and the concentration of cell factors can be kept by the 3D cell culture material for a long time, and accordingly the 3D cell culture material is favorable for fast growing and proliferating cells and maintaining the activity of the cells; the cell factors can be conveniently and easily fixed by the aid of nano-particles NH2@Fe<3>O<4> in the 3D cell culture material, and proteins of the cell factors can be combined with external residues of micelle by the aid of hydrogen bonds under the condition of low-concentration glutaraldehyde; the problem of low culture efficiency due to dissociation of proteins in water bases can be solved; excellent suppression and culture effects for tumor cells and stem cell differentiation can be realized by the 3D cell culture material; culture requirements of different primary mammal cells can be met.

Description

technical field [0001] The invention belongs to the technical field of material chemistry and biomedicine, and in particular relates to a material for three-dimensional culture of cells in vitro and its application. Background technique [0002] Three-dimensional cell culture technology (three-dimensional cell culture, TDCC) refers to the co-cultivation of carriers with different materials with three-dimensional structures and various types of cells in vitro, so that cells can migrate and grow in the three-dimensional space structure of the carrier, forming Three-dimensional cell-carrier complex. [0003] Three-dimensional cell culture technology has important applications. Ordinary cell culture gradually loses its original traits due to the proliferation of cells in a changed environment in vitro, which is often inconsistent with the situation in the body. Animal experiments are completely carried out in vivo, but due to various factors in the body and the interaction betw...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12N5/0775C12N5/07
Inventor 邹寰陈杰曾宏彬余旻斐张璐
Owner GUANGZHOU SAGENE BIOTECH
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