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Three-dimensional space cell culture system preparation method

A cell culture, three-dimensional space technology, applied in the field of cell biology and tissue engineering, can solve the problems of non-permeability of scaffold materials, unfavorable cell planting treatment and tissue transplantation or implantation, cytotoxicity, etc.

Active Publication Date: 2012-07-18
杨炜 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A major limitation of collagen-based scaffolding techniques is that cells tend to grow on the surface of collagen scaffolds due to lack of nutrients for cells located deep within the scaffold
For example, when Matrigel is used as the extracellular matrix, when the cultured cells are separated from this matrix, enzymes are used to degrade or dissolve the gel, which can cause cell damage
Some matrices lack good biocompatibility, which may cause cytotoxicity, or teratogenic or tumorigenic effects
Some matrices have poor biodegradability, which is not conducive to cell seeding therapy and tissue transplantation or implantation
In addition, most scaffold materials are not transparent, and it is difficult to directly observe the cultured tissue cells at any time and dynamically, especially under the condition of not destroying the "specimen"
In addition, the currently used three-dimensional space culture system has not been able to obtain the cultured cells that can maintain long-term cell proliferation
The number of cells produced by the existing method is not enough, the size or thickness of the tissue is not enough, it lacks the strength it should have, and it cannot mature, differentiate and grow into a shape similar to the peculiar shape of normal tissues and organs in the body

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0183] Example 1 The structure of the see-through three-dimensional cell culture unit

[0184] In this embodiment, agar is selected as the material for making the see-through three-dimensional cell culture unit.

[0185] The production mold of the see-through three-dimensional cell culture unit is as follows: figure 1 and figure 2 As shown, first design a cuboid metal jacket, and place 3 square metal wires in the jacket.

[0186] In this embodiment, the size of the rectangular jacket is designed as follows: the length is 10mm, the width is 6.5mm, the height is 3mm, and the wall thickness is 0.5mm.

[0187] The length of the square metal wire is 20mm, and the width and height are respectively 0.9mm.

[0188] Both ends of the outer sleeve are sealed by removable caps matching the outer sleeve.

[0189] Dissolve 1% agar in PBS solution of pH 7.4, disinfect and inject it into the jacket. After the agar-injected outer tube was placed at room temperature for 15 minutes, the ...

Embodiment 2

[0192] Example 2 Comparison of cells implanted into different three-dimensional space cell culture units of the present invention

[0193] In this example, the method described in Example 1 was used to prepare a three-dimensional cell culture unit with a circular cross section and a circular inner cavity. The outer diameter of the mold with a circular cross section is about 6 mm, the inner diameter is about 5 mm, the wall thickness is about 0.5-1 mm, and the length is 200 mm. The diameter of the circular metal wire is 0.2 mm, and the length is 300 mm. 5-8 metal wires can be placed in the mold, and agar gel is added into the mold to prepare a three-dimensional cell culture unit with a circular inner cavity cross section.

[0194] The results of cell seeding into the three-dimensional cell culture units with square and circular cross-sections prepared above are as follows: image 3 shown. from image 3 It can be seen in the figure that the inner cavity is a square three-dim...

Embodiment 3

[0195] Example 3 Culture of liver cancer cell line SMMC 7721

[0196] In this example, the liver cancer cell line SMMC 7721 was cultured using the three-dimensional cell culture unit with a circular cross section prepared by the method in Example 2. The cell culture medium used is RPMI 1640 (Sigma R 6504) 1000ml, 100 times penicillin + streptomycin (GIBCO 15140-122) 10ml, fetal bovine serum 100ml. The culture conditions were 5% carbon dioxide and the temperature was 37°C.

[0197] In addition, the inventors also cultured liver cancer cell line SMMC 7721 in a general two-dimensional culture system (ordinary cell culture dish) under the same medium, culture temperature and other conditions as a control.

[0198] The situation of the growth of liver cancer cell line SMMC 7721 in two-dimensional and the culture system of the present invention is as follows: Figure 4 shown, where Figure 4 A (10X) is the culture and growth of liver cancer cell line SMMC 7721 in the traditiona...

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Abstract

The invention provides a three-dimensional space cell culture system preparation method. A fine chain is placed in an outer casing pipe, biological degradable materials are melted and then filled in the outer casing pipe, and after the biological degradable materials are solidified and sized, the outer casing pipe and the fine chain are removed, so that a three-dimensional cell culture unit is prepared. The three-dimensional space culture unit prepared by the method can be applied in long-period cell culturing and a large amount of cells generating, the large amount of cells serve as seed cells of tissue engineering, and the three-dimensional space culture unit can be applied in transplant treatment by generating regeneration tissue and micro-organs in a short time.

Description

[0001] The present invention is a divisional application of the following application. [0002] Invention name: Visible three-dimensional space cell culture system and its application in the culture of tissue cells and neonatal organs [0003] Application date: 2006.1.23 [0004] Application number: 200610023537.5. technical field [0005] The invention belongs to the fields of cell biology and tissue engineering, and in particular relates to a method for preparing a three-dimensional space cell culture system and its application in the culture of tissue cells and neonatal organs. Background technique [0006] Tissue cell culture is an important method for studying normal, diseased and malignant cells. Tissue and cell culture techniques can be used to observe, set or change the whole process of activation, proliferation, differentiation, migration, maturation, aging, shedding and even cell death of different types of cells and their regulatory mechanisms under specific con...

Claims

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Application Information

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IPC IPC(8): C12M3/00
Inventor 杨炜刘华
Owner 杨炜
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