3D (three-dimensional) cell culture material and application thereof

A technology of cells and culture medium, which is applied to the materials of three-dimensional culture of cells in vitro and their application fields, to achieve the effect of changing their own survival and development, changing conditions, and maintaining their own survival and development.

Active Publication Date: 2015-08-05
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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  • 3D (three-dimensional) cell culture material and application thereof
  • 3D (three-dimensional) cell culture material and application thereof

Examples

Experimental program
Comparison scheme
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, so that 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 and 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 100~200n...

Embodiment

[0092] Example 3 Application of a three-dimensional cell culture material

[0093] Get the PEG8000Fe prepared by embodiment 1 respectively 3 o 4 25mg, PEG400 Fe 3 o 4 Add 10mg to 10mL GIBICO's DMEM Basal Medium medium, shake and mix well, and then pass through 300W / m 2 Ultrasound with sound intensity is used for drilling for 3-5 minutes to form a uniform three-dimensional cavity, and can produce holes of the same size as the microenvironment, which is convenient for direct mixing and planting cells; finally, human embryonic kidney 293T cells are inoculated into the culture medium , mixed with weak ultrasonic vibration 1-2mim.

[0094] 3) Cultivation: The cell culture in the previous step was cultured with temperature-controlled stirring using a magnetic stirrer for mammalian fermentation, and other culture conditions were the same as those indicated in the existing recipe.

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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 structure and various types of cells in vitro, so that cells can migrate and grow in the three-dimensional space structure of the carrier to form Three-dimensional cell-carrier complex. [0003] Three-dimensional cell culture technology has important applications. Ordinary cell culture is often inconsistent with the situation in vivo because cells proliferate and gradually lose their original traits in the changed environment in vitro, while animal experiments are completely carried out in vivo, but due to various factors in the body and the interaction between the ...

Claims

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Application Information

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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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