Permeable and visible tri-dimensional cell culture system and its uses in tissue and newborn organ culture

A three-dimensional cell, cell technology, applied in the field of cell biology and tissue engineering, can solve the problems of lack of biocompatibility, lack of transparency of scaffold materials, insufficient number of cells, etc.

Active Publication Date: 2007-08-01
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, mo...

Method used

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  • Permeable and visible tri-dimensional cell culture system and its uses in tissue and newborn organ culture
  • Permeable and visible tri-dimensional cell culture system and its uses in tissue and newborn organ culture
  • Permeable and visible tri-dimensional cell culture system and its uses in tissue and newborn organ culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0179] Example 1 The structure of the see-through three-dimensional cell culture unit

[0180] In this embodiment, agar is selected as the material for making the see-through three-dimensional cell culture unit.

[0181] The production mold of the see-through three-dimensional cell culture unit is shown in Figure 1 and Figure 2. First, a cuboid metal jacket is designed, and three square metal wires are placed in the jacket.

[0182] 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.

[0183] The length of the square metal wire is 20mm, and the width and height are respectively 0.9mm.

[0184] Both ends of the outer sleeve are sealed by removable caps matching the outer sleeve.

[0185] Dissolve 1% agar in PBS solution of pH 7.4, disinfect and inject into the jacket after dissolving. After the agar-injected outer tube was placed at room temperature for 15 m...

Embodiment 2

[0188] Example 2 Comparison of cells implanted into different three-dimensional space cell culture units of the present invention

[0189] 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.

[0190] The results of cell implantation into the three-dimensional cell culture units with square and circular cross-sections prepared above are shown in FIG. 3 . It can be seen from Figure 3 that the three-dimensional cell culture unit with a square inner c...

Embodiment 3

[0191] Example 3 Culture of liver cancer cell line SMMC 7721

[0192] 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 was 1000ml of RPMI 1640 (Sigma R6504), 10ml of 100-fold penicillin+streptomycin (GIBCO 15140-122), and 100ml of fetal bovine serum. The culture conditions were 5% carbon dioxide and a temperature of 37°C.

[0193] 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.

[0194] The situation of the growth of liver cancer cell line SMMC 7721 in two-dimensional and culture system of the present invention is as shown in Figure 4, wherein Fig. 4A (10X) is the culture growth situation of liver cancer cell line SMMC 7721 in traditional two-di...

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Abstract

The invention discloses a three-dimensional cell culture system, comprising container, and cell culturing medium and three- dimentioal cell culture unit inside container; said three- dimentional cell culture unit comprises implanting cell and cavity flavoable for cell adsoption, growth, differentiation and maturity. Said three- dimentional culture unit is transparent, which is flavouable for obeservation of adsorption, enlargement, transition, proliferation, differentiation, maturity, aging, death and tissue or organ development. The invention can be used for long term cell culture and large amount of cell production as seed cell for tissue engineering, and for short term regenerative tissue and micro organ formation for replanting. The invention provides special condition for tumor external research, and convenient for tumor diagnosis, observation of tumor immersion and transfer, and anti- tumor medicine sift. It can aslo be used for cell isolated culture and for research of gene programm change and influence to cell from rxternal environment.

Description

technical field [0001] The invention belongs to the fields of cell biology and tissue engineering, and in particular relates to a three-dimensional space cell culture system and its application in the culture of tissue cells and neonatal organs. Background technique [0002] 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 conditions in vitro. In the past 50 years, tissue cell culture has played an important role in promoting the development of basic medical science research and the improvement of clinical treatment. [0003] A variety of diseases or trauma seen clinically can cause cell degeneration, necrosis or damage, which in turn leads to damage or loss of t...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12N5/08C12N5/00C12N5/071
Inventor 杨炜刘华
Owner 杨炜
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