CN103255058A - Simulation three-dimensional incubator and culture method for mouth mucosa epithelial cells and applications of incubator and culture method - Google Patents
Simulation three-dimensional incubator and culture method for mouth mucosa epithelial cells and applications of incubator and culture method Download PDFInfo
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- CN103255058A CN103255058A CN2013101710910A CN201310171091A CN103255058A CN 103255058 A CN103255058 A CN 103255058A CN 2013101710910 A CN2013101710910 A CN 2013101710910A CN 201310171091 A CN201310171091 A CN 201310171091A CN 103255058 A CN103255058 A CN 103255058A
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Abstract
The invention discloses a simulation three-dimensional incubator and a culture method for mouth mucosa epithelial cells and applications of the incubator and the culture method. The incubator comprises a single or a plurality of basic structural units, wherein each basic structural unit consists of an inner container and an outer container which are tightly nested with each other, and are prepared from an organic material for cell culture; and the bottom of the inner container consists of a permeable supporting film, and a certain gap is formed between the bottom of the inner container and the bottom of the outer container. The incubator takes the factors of nutrient supply of the mouth mucosa epithelial cells, blood vessels and blood vessel endothelial cells into account, and the growing environment of the mouth mucosa epithelial cells in the incubator is close to that in a human body, so that the incubator can be used for physiological, pharmacological, toxicological and micro-ecological studies of mouth mucosas, and can also be used for constructing human tissue engineering mouth mucosas and repairing and reconstructing mucosal coloboma. The new technology of the simulation three-dimensional incubator promotes an in vitro culture model of a mouth mucosa to a new height, so that related studies are closer to clinical practices.
Description
Technical field
The present invention relates to field of cell culture, be specifically related to a kind of oral mucosa epithelial cell emulation dimensional culture device and cultural method and application.
Background technology
The oral mucosa epithelial cell vitro culture has important biology and clinical meaning.It not only provides good experimental system for the differentiation of research oral mucosa, oral cavity pathologic, physiologic, toxicology and oncogenesis mechanism etc., also significant to human tissue through engineering approaches oral mucosa structure and oral mucosas tissue's defect repair, the reconstruction for mucomembranous defects such as eyelid, larynx, tracheae and urethras has important clinical significance simultaneously.
Cells whose development and function all depend on intermolecular interaction in the cell micro-environment.Traditional two-dimentional cell cultures is comparatively general cell cultures means, be not the state of nature of cell growth, thereby cell all may there are differences aspect genetic expression, signal transduction and the morphology with native state.The three-dimensional cell cultivation is the experimental technique between experimentation on animals and two-dimensional flat plate cell cultures, and three-dimensional carrier and cell are cultivated altogether external, makes cell move in the solid space structure of carrier, grow, and constitutes three-dimensional cell carrier mixture.The dimensional culture system provides support or the matrix of growing environment in the analog for cell, cell by closely connect with (or) slit mode such as is connected and sets up contacting between iuntercellular and cell and matrix, forms certain three-dimensional structure; Three-dimensional cell is cultivated at aspects such as genetic expression, matrix secretion and cell function activities all has notable difference with the two dimension cultivation, more similar to the cells in vivo growing environment.Therefore, three-dimensional cell is cultivated the three-dimensional microenvironment of analogue body inner cell growth more realistically, and the response of cell under three-dimensional condition more can represent its response under condition in vivo, can embody intuitive and the condition controllability of cell cultures again.
At present, the dimensional culture technology has been applied in the in-vitro culture model of kinds of tumor cells and stem cell etc., but does not see the dimensional culture device of oral mucosa epithelial cell and the report of cultural method.
Summary of the invention
The objective of the invention is to the deficiency that exists according in the existing oral mucosa epithelial cell culture technique, a kind of oral mucosa epithelial cell emulation dimensional culture device is provided, highly to approach the physiological environment of simulation human body oral mucosa;
Another object of the present invention is to provide a kind of cultural method that uses above-mentioned oral mucosa epithelial cell emulation dimensional culture device;
A further object of the invention is to provide the application of above-mentioned oral mucosa epithelial cell emulation dimensional culture device.
For realizing the object of the invention, the scheme that the present invention takes is: a kind of oral mucosa epithelial cell emulation dimensional culture device, it comprises single or multiple basic structural units, described basic structural unit is made up of tight nested inner jar and outer container, and described inner jar and outer container are made by the organic materials that is used for cell cultures; The bottom of inner jar is made of the infiltration supporting film and has certain interval with the bottom of outer container.
Further preferred scheme, described inner jar and outer container are cylindric.
Further preferred scheme, described infiltration supporting film is made by the organic materials that is used for cell cultures, and the infiltration supporting film is provided with intensive aperture, is used for nutrient solution or other composition and passes through; Hole diameter is much smaller than cell dia.
In addition, the invention provides a kind of cultural method of oral mucosa epithelial cell emulation dimensional culture device, it may further comprise the steps:
Vascular endothelial cell is inoculated in and forms the vascular endothelial cell layer on the infiltration supporting film of inner jar; Mesenchymal cell mixes with extracellular matrix and is positioned over infiltration supporting film vascular endothelial cell layer top; In the top of mixture inoculation oral mucosa epithelial cell; Fresh medium or the fresh medium that contains certain concentration medicine are added in the outer container, see through the vascular endothelial cell layer and enter extracellular matrix contact oral mucosa epithelial cell, provide nutritive ingredient or medicine to oral mucosa epithelial cell.
Further preferred scheme, described mesenchymal cell comprises fibrocyte and immunocyte.
Further preferred scheme according to the length of incubation time, determines to add the amount of nutrient solution, and the periodic replacement nutrient solution.
The scheme that the present invention substitutes, drugs with function also can directly add from the top and act on oral mucosa epithelial cell, with the direct administering mode of simulation human body oral mucosa.
Also have, the invention provides a kind of described oral mucosa epithelial cell emulation dimensional culture device and the application of cultural method in physiology, pharmacology, toxicology and little ecological Studies of oral mucosa.
Also have, the invention provides the application in the reconstruction of human tissue through engineering approaches oral mucosa structure and mucomembranous defect of a kind of described oral mucosa epithelial cell emulation dimensional culture device and cultural method.
Compared with prior art, the present invention has considered the factor of nutrition supply, blood vessel and the vascular endothelial cell of oral mucosa epithelial cell, close to the growing environment of human oral cavity mucous membrane, makes research more near clinical practice.
Description of drawings
Fig. 1 is the structural representation of oral mucosa epithelial cell emulation dimensional culture device;
Fig. 2 is the structural representation of inner jar;
Fig. 3 is the structural representation of outer container.
Embodiment
Further explain the present invention below in conjunction with embodiment, but embodiment does not do any type of restriction to the present invention.
As shown in Figure 1 to Figure 3, a kind of oral mucosa epithelial cell emulation dimensional culture device, it comprises single or multiple basic structural units, and described basic structural unit is made up of tight nested inner jar 1 and outer container 2, and described inner jar 1 and outer container 2 are made by the organic materials that is used for cell cultures; The bottom of inner jar 1 is made of infiltration supporting film 3 and has certain interval with the bottom of outer container 2.This device can be constituted or is made of inner jar, outer container that a series of one-tenth are organized row in length and breadth by single inner jar, outer container.
Further preferred scheme, described inner jar 1 and outer container 2 are cylindric.
Further preferred scheme, described infiltration supporting film 3 is made by the organic materials that is used for cell cultures, and infiltration supporting film 3 is provided with intensive aperture, is used for nutrient solution or other composition and passes through; Hole diameter is much smaller than cell dia, so vascular endothelial cell 4 can not drop and can form the vascular endothelial cell layer at film.
In addition, the invention provides a kind of cultural method of oral mucosa epithelial cell emulation dimensional culture device, it may further comprise the steps:
Vascular endothelial cell 4 is inoculated in and forms the vascular endothelial cell layer on the infiltration supporting film 3 of inner jar 1; Mesenchymal cell 6 mixes with extracellular matrix 5 and is positioned over infiltration supporting film 3 vascular endothelial cell layers 4 top; In the top of mixture inoculation oral mucosa epithelial cell 7; Fresh medium 8 or the fresh medium that contains certain concentration medicine are added in the outer container 2, see through vascular endothelial cell layer 4 and enter extracellular matrix 5, mesenchymal cell 6 contact oral mucosa epithelial cells 7, provide nutritive ingredient or medicine to oral mucosa epithelial cell 7.
Further, described mesenchymal cell 6 comprises fibrocyte and immunocyte.Separate in the unnecessary oral mucosa that oral mucosa epithelial cell 7 and various mesenchymal cells 6 can abandon from hospital's decorative sursery operation and obtain or buy from commercially available cell strain.
Further, according to the length of incubation time, determine to add the amount of nutrient solution, and the periodic replacement nutrient solution.
Except aforesaid method, drugs with function also can directly add from the top and acts on oral mucosa epithelial cell 7, with the direct administering mode of simulation human body oral mucosa, thereby carries out physiology, pharmacology, toxicology and little ecological Studies.
Also have, the invention provides a kind of described oral mucosa epithelial cell emulation dimensional culture device and the application of cultural method in physiology, pharmacology, toxicology and little ecological Studies of oral mucosa.
Also have, the invention provides the application in the reconstruction of human tissue through engineering approaches oral mucosa structure and mucomembranous defect of a kind of described oral mucosa epithelial cell emulation dimensional culture device and cultural method.
Claims (9)
1. oral mucosa epithelial cell emulation dimensional culture device, it is characterized in that, comprise single or multiple basic structural units, described basic structural unit is made up of tight nested inner jar and outer container, and described inner jar and outer container are made by the organic materials that is used for cell cultures; The bottom of inner jar is made of the infiltration supporting film and has certain interval with the bottom of outer container.
2. according to the described oral mucosa epithelial cell emulation of claim 1 dimensional culture device, it is characterized in that described inner jar and outer container are cylindric.
3. according to the described oral mucosa epithelial cell emulation of claim 1 dimensional culture device, it is characterized in that described infiltration supporting film is made by the organic materials that is used for cell cultures, the infiltration supporting film is provided with intensive aperture, is used for nutrient solution or other composition and passes through; Hole diameter is much smaller than cell dia.
4. the cultural method according to each described oral mucosa epithelial cell emulation dimensional culture device in the claim 1 ~ 3 is characterized in that, may further comprise the steps:
Vascular endothelial cell is inoculated in and forms the vascular endothelial cell layer on the infiltration supporting film of inner jar; Mesenchymal cell mixes with extracellular matrix and is positioned over infiltration supporting film vascular endothelial cell layer top; In the top of mixture inoculation oral mucosa epithelial cell; Fresh medium or the fresh medium that contains certain concentration medicine are added in the outer container, see through the vascular endothelial cell layer and enter extracellular matrix contact oral mucosa epithelial cell, provide nutritive ingredient or medicine to oral mucosa epithelial cell.
5. cultural method according to claim 4 is characterized in that, described mesenchymal cell comprises fibrocyte and immunocyte.
6. cultural method according to claim 4 is characterized in that, according to the length of incubation time, determines to add the amount of nutrient solution, and the periodic replacement nutrient solution.
7. cultural method according to claim 4 is characterized in that, drugs with function also can directly add from the top and acts on oral mucosa epithelial cell, with the direct administering mode of simulation human body oral mucosa.
8. one kind according to each described oral mucosa epithelial cell emulation dimensional culture device in the claim 1 ~ 7 and the application of cultural method in physiology, pharmacology, toxicology and little ecological Studies of oral mucosa.
One kind according to each described oral mucosa epithelial cell emulation dimensional culture device and cultural method in the claim 1 ~ 7 human tissue through engineering approaches oral mucosa make up and the reconstruction of mucomembranous defect in application.
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| CN2013101710910A CN103255058A (en) | 2013-05-10 | 2013-05-10 | Simulation three-dimensional incubator and culture method for mouth mucosa epithelial cells and applications of incubator and culture method |
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| CN2013101710910A CN103255058A (en) | 2013-05-10 | 2013-05-10 | Simulation three-dimensional incubator and culture method for mouth mucosa epithelial cells and applications of incubator and culture method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110072988A (en) * | 2016-10-28 | 2019-07-30 | 国立大学法人京都大学 | The co-culture device and co-culture method of the bacteriums such as Anaerobic Bacteria and epithelial cell |
| CN111665304A (en) * | 2020-06-02 | 2020-09-15 | 南京融康博生物科技有限公司 | Tumor cell culture information analysis method |
Citations (2)
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| US20060153815A1 (en) * | 2004-12-21 | 2006-07-13 | Agnieszka Seyda | Tissue engineering devices for the repair and regeneration of tissue |
| CN102399693A (en) * | 2011-10-25 | 2012-04-04 | 广东工业大学 | Simulation three-dimensional cell cultivation device and cultivation method |
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2013
- 2013-05-10 CN CN2013101710910A patent/CN103255058A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060153815A1 (en) * | 2004-12-21 | 2006-07-13 | Agnieszka Seyda | Tissue engineering devices for the repair and regeneration of tissue |
| CN102399693A (en) * | 2011-10-25 | 2012-04-04 | 广东工业大学 | Simulation three-dimensional cell cultivation device and cultivation method |
Non-Patent Citations (2)
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| 于世凤主编: "《口腔组织病理学》", 31 January 2001 * |
| 魏昕等: "口腔黏膜上皮细胞和成纤维细胞的复合培养", 《临床口腔医学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110072988A (en) * | 2016-10-28 | 2019-07-30 | 国立大学法人京都大学 | The co-culture device and co-culture method of the bacteriums such as Anaerobic Bacteria and epithelial cell |
| CN111665304A (en) * | 2020-06-02 | 2020-09-15 | 南京融康博生物科技有限公司 | Tumor cell culture information analysis method |
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Application publication date: 20130821 |