CN112210498A

CN112210498A - Cartilage diaphragm autocrine factor retention culture dish

Info

Publication number: CN112210498A
Application number: CN202011123239.XA
Authority: CN
Inventors: 白宝帅; 周广东; 侯梦洁; 徐伟; 郝俊祥; 刘豫; 曹谊林
Original assignee: Shanghai Soft Heart Biotechnology Co ltd
Current assignee: Shanghai Soft Heart Biotechnology Co ltd
Priority date: 2020-10-20
Filing date: 2020-10-20
Publication date: 2021-01-12

Abstract

The invention discloses a culture dish for retaining autocrine factors of a cartilage membrane, which structurally comprises a culture body and a semi-permeable membrane body, wherein the culture body is connected with the semi-permeable membrane body in a clamping manner, a certain distance is arranged between the bottom of the membrane cavity at the bottom of the semi-permeable membrane body and the bottom of a dish at the bottom of the culture body, and the connecting edge of the culture body and the semi-permeable membrane body is designed to be clamped by a stepped bayonet; the semi-permeable membrane body consists of a supporting member and a semi-permeable membrane, wherein the semi-permeable membrane is a fiber filtering layer wrapping the supporting member and divides the whole cavity into a space body above the membrane and a space body below the membrane; a space body under the membrane and the bottom of the square dish are used as a cell planting space body; according to the invention, cells are planted in a cell planting space body, a corresponding culture medium and factors are added, then a semi-permeable membrane body is installed, water molecules and harmful molecules can slowly reach the space body on the membrane through an osmotic gradient, and the purpose of changing liquid is achieved by changing the liquid of the space body on the membrane every day; meanwhile, the invention has reasonable and simple structural design, is easy to realize and is suitable for popularization and application.

Description

Cartilage diaphragm autocrine factor retention culture dish

Technical Field

The invention relates to the technical field of cartilage membrane culture dishes, in particular to a cartilage membrane autocrine factor retention culture dish.

Background

Articular cartilage is mainly a vascular-free tissue composed of chondrocytes and extracellular matrix, and mainly depends on the movement and extrusion of joints to absorb nutrients, so that the self-repair capacity of cartilage after damage is weak, the clinical research has limited material for patient tissues, and the chondrocytes are terminally differentiated cells, have limited in vitro proliferation capacity and are easy to dedifferentiate, wherein the dedifferentiation refers to the process that differentiated cells lose specific structures and functions and are changed into undifferentiated cells. Therefore, the improvement and maintenance of the phenotype of the chondrocytes and the promotion of chondroiysis of the chondrocytes play important roles in clinical application of the chondrocytes.

In the prior art, high-density chondrocytes are inoculated in a culture well plate, and the chondrocytes are cultured by chondrogenic induction liquid for 2 weeks, during which the culture medium is replaced 1 or 2 times per day; the culture medium stimulates the chondrocytes to secrete cartilage extracellular matrix, and the cartilage extracellular matrix wraps the chondrocytes to obtain the cartilage cell membrane, wherein the cartilage extracellular matrix mainly comprises type ii collagen and glycosaminoglycan.

In addition, an improved culture medium is added from an upper end cover above a cell culture dish container body, the cell suspension is added from an arc-shaped cell suspension adding port arranged on a side wall, a polycarbonate semi-permeable membrane capable of filtering molecules with the molecular weight of 5kb is arranged between the cell suspension and the improved culture medium, the semi-permeable membrane allows metabolic waste and nutrient components to freely pass through, but can not allow factors promoting chondrogenesis to pass through, so that liquid above the semi-permeable membrane is only changed every time liquid is changed, chondrogenesis promoting factors do not need to be additionally added, autocrine factors generated by cells are protected, the production cost is reduced, the product can contain more liquid, the operation of changing liquid every day is avoided, the pollution possibility and heavy work are reduced, the cartilage cell membrane culture is realized by using a traditional pore plate or a culture dish, and the cell amount is very high, so that expensive chondrogenesis induced liquid needs to be changed every day, high cost and the need to perform daily operations increases the likelihood of contamination of the cartilage membrane and the workload of the laboratory personnel.

In order to promote market selectivity and competitiveness, the skilled person should therefore also address the above issues in time.

Disclosure of Invention

The invention aims to solve the problems and provide a cartilage membrane autocrine factor retention culture dish, which can solve the problems of high cost of a cartilage formation induction process and pollution and heavy workload caused by liquid change through simple structural design.

In order to achieve the above purpose, the technical scheme of the invention is as follows: the utility model provides a culture dish is remained to cartilage diaphragm autocrine factor, its characterized in that, the structure includes culture body and semi-permeable membrane body two parts, and culture body and semi-permeable membrane body chucking are connected, are provided with the certain distance between the bottom of semi-permeable membrane body bottom membrane chamber and the culture body bottom side dish, and culture body and semi-permeable membrane body connecting edge department design are step bayonet socket chucking.

Preferably, the semipermeable membrane body is composed of a support member and a semipermeable membrane, the semipermeable membrane is a fiber filtration layer wrapping the support member, and the whole cavity is divided into a space above the membrane and a space below the membrane.

Preferably, the space under the membrane and the bottom of the dish are used as a cell planting space.

Preferably, the semipermeable membrane can carry molecules with molecular weights of 300, 500, 1000, 3500, 7000 and 14000 according to different models.

According to the invention, cells are planted in a cell planting space body, a corresponding culture medium and factors are added, then a semi-permeable membrane body is installed, water molecules and harmful molecules can slowly reach a space body on the membrane through an osmotic gradient, and the purpose of changing liquid is achieved by changing liquid of the space body on the membrane; the culture dish can retain factors of the original chondrogenic culture medium and cell autocrine factors, has important effect on reconstructing a cartilage microenvironment by in vitro chondrocytes and promotes the in vitro growth of the cartilage; the culture dish can be used for culturing cartilage membranes, plays a very important role in culturing other types of cells, protects autocrine factors generated by the cells, reduces the production cost, reduces the pollution possibility and heavy work, and is reasonable and simple in structural design, easy to implement and suitable for popularization and application.

Drawings

FIG. 1 is a schematic diagram of the structural relationship of the present invention;

the labels in the figure are: 1-a culture; 11-square dish bottom; 2-a semi-permeable membrane body; 21-membrane cavity bottom; 22-a support member; 23-a semi-permeable membrane; 24-intermediates on membranes; 25-subintimal space volume; 3-step type bayonet; 4-cell implant space.

Detailed Description

The present invention will be described in detail below with reference to the accompanying drawings.

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

As shown in the figure, a culture dish is remained to cartilage diaphragm autocrine factor, its characterized in that, the structure includes culture body 1 and semi-permeable membrane body 2 two parts, and culture body 1 and semi-permeable membrane body 2 chucking are connected, are provided with the certain distance between 2 bottom membrane chamber bottoms 21 of semi-permeable membrane body and the culture body 1 bottom side dish bottom 11, and culture body 1 and semi-permeable membrane body 2 connection edge department design is 3 chuckings of step bayonet socket.

In the specific implementation process, the semi-permeable membrane body 2 is composed of a supporting member 22 and a semi-permeable membrane 23, wherein the semi-permeable membrane 23 is a fiber filtering layer wrapping the supporting member 22 and divides the whole cavity into a membrane upper space 24 and a membrane lower space 25.

In the specific implementation, the space under the membrane 25 and the bottom of the square dish 11 serve as the cell planting space 4.

In the specific implementation process, the semipermeable membrane can carry and retain molecules with molecular weights of 300, 500, 1000, 3500, 7000 and 14000 according to different models.

In the specific implementation process, the cells are planted in the cell planting space body 4 in the step 1, corresponding culture media and factors are added, the semi-permeable membrane body 2 is installed in the step 2, water molecules and harmful molecules can slowly reach the space body 24 on the membrane through osmotic gradient, and the purpose of liquid change is achieved by replacing liquid of the space body 24 on the membrane; the culture dish can retain factors of the original chondrogenic culture medium and cell autocrine factors, has important effect on reconstructing a cartilage microenvironment by in vitro chondrocytes and promotes the in vitro growth of the cartilage; the culture dish can be used for culturing cartilage membranes, plays a very important role in culturing other types of cells, protects autocrine factors generated by the cells, reduces the production cost, reduces the pollution possibility and heavy work, and is reasonable and simple in structural design, easy to implement and suitable for popularization and application.

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.

Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.

Claims

1. The utility model provides a culture dish is remained to cartilage diaphragm autocrine factor, its characterized in that, the structure includes culture body and semi-permeable membrane body two parts, and culture body and semi-permeable membrane body chucking are connected, are provided with the certain distance between the bottom of semi-permeable membrane body bottom membrane chamber and the culture body bottom side dish, and culture body and semi-permeable membrane body connecting edge department design are step bayonet socket chucking.

2. The cartilage membrane autocrine factor retaining culture dish of claim 1, wherein the semi-permeable membrane body is composed of a supporting member and a semi-permeable membrane, the semi-permeable membrane is a fiber filtering layer wrapping the supporting member to divide the whole cavity into a supramembrane space and a sub-membrane space.

3. The cartilage membrane autocrine factor retention culture dish of any of claims 1 or 2, wherein the cell seeding space is between the membrane inferior space and the bottom of the square dish.

4. The cartilage membrane autocrine factor retaining culture dish of claim 2, wherein the semi-permeable membrane can carry molecules with molecular weight of 300, 500, 1000, 3500, 7000 and 14000 according to different types.

5. The cartilage membrane autocrine factor retention culture dish of any one of claims 1-4, wherein step 1 is to plant cells in a cell planting space and add the corresponding culture medium and factor; and 2, installing the semi-permeable membrane body, wherein water molecules and harmful molecules can slowly reach the space body on the membrane through osmotic gradient, and replacing the liquid in the space body on the membrane to achieve the purpose of replacing the liquid.