CN110724637A - Glass slide system for direct contact cell co-culture and working method thereof - Google Patents

Abstract

The invention provides a slide glass system for direct contact cell co-culture, which comprises a slide glass culture box, a cell culture slide glass arranged on the slide glass culture box, and a separating frame which is detachably arranged on the cell culture slide glass; the slide system also includes a separator for separating the spacer from the cell culture slides. The invention also provides a working method of the slide glass system for direct contact cell co-culture, which comprises the following steps: (1) placing a cell culture slide in a slide culture box; (2) fixing the separation frame on a cell culture glass slide; (3) after the cells adhere to the wall, the separating frame is removed, and the glass slide is still placed in a glass slide culture box for culture; (4) the cells interact with each other. The slide glass system for direct contact cell co-culture has the advantages of simple structure, convenient use, simple operation, less required culture solution, economy and environmental protection.

Description

Glass slide system for direct contact cell co-culture and working method thereof

Technical Field

The invention belongs to the technical field of cell culture, and particularly relates to a glass slide system for co-culture of direct contact cells and a working method thereof.

Background

All organisms are organic whole bodies composed of a plurality of cells, and different cells can maintain and regulate various activities of the organisms through interaction and mutual influence. For example: cellular communication between neurons and oligodendrocytes affects myelin formation, the interaction of macrophages and immunocompetent cells is involved in antigen extraction processes in immune responses, the interaction between astrocytes and fibroblasts is involved in glial scar formation, and so on. The interaction and interaction between the cells are very extensive and ubiquitous, and the research on the systematicness is worth.

Cell-cell interactions are commonly studied by co-culturing different types of cells. Currently, there are two main ways: non-direct contact co-culture and direct contact co-culture. The non-direct contact type co-culture is that different types of cells to be researched are respectively inoculated to different carriers and then are placed in the same culture system, but the cells are not contacted with each other, and the non-direct contact type co-culture is only suitable for the research of the interaction of the cells through paracrine cell factors and the like; the direct contact type co-culture is to culture different types of cells in the same culture system, but the direct contact mixed culture of different types of cells has the most obvious defect that the cells are mixed together and are not beneficial to subsequent observation, detection and analysis. In fact, this puts new demands on direct contact co-culture, and there is no model for a direct co-culture system of two or more cells that is both simple and meets the new demands and facilitates subsequent observation and detection.

Disclosure of Invention

The invention provides a direct-contact cell co-culture glass slide system and a working method thereof, which aim to solve the problems in the background art.

In order to solve the above technical problem, an embodiment of the present invention provides a slide glass system for direct contact cell co-culture, which is characterized in that the slide glass system comprises a slide glass culture box, a cell culture slide placed in the slide glass culture box, and a separation frame detachably disposed on the cell culture slide; the slide system also includes a separator for separating the spacer from the cell culture slides.

Furthermore, the slide glass culture box comprises a box body and a cover body which is arranged above the box body in a separable mode, and the size of the cover body is matched with that of the box body.

Preferably, one side of the cell culture glass slide is provided with a single-side frosted surface, and the upper end face of the cell culture glass slide is coated with polylysine.

Furthermore, the lower end face of the separation frame is provided with a biocompatible acrylic adhesive, and the separation frame is hermetically connected with the cell culture glass slide through a viscous substance.

Furthermore, the cell culture glass slide is made of white borosilicate glass, and the cell culture glass slide is 25mm multiplied by 75mm in size and 1-1.2mm in thickness.

Furthermore, the separation frame is made of plastic materials.

Furthermore, the separating frame is divided into an outer frame and an inner separating plate, the thickness of the outer frame is 1mm, the thickness of the inner separating plate is 0.1mm, 0.3mm or 0.5mm, and the heights of the outer frame and the inner separating plate are 10 mm.

Further, the separating frame can be divided into a four-hole frame, a six-hole frame and an eight-hole frame, the size of a hole in the four-hole frame is 12mm multiplied by 23mm, the size of a hole in the six-hole frame is 8mm multiplied by 23mm, and the size of a hole in the eight-hole frame is 12mm multiplied by 11.35 mm.

Furthermore, the slide glass culture box is made of transparent polystyrene, and the size of the slide glass culture box is 83mm multiplied by 30mm multiplied by 15 mm.

The embodiment of the invention also provides a working method of the slide system for the direct contact cell co-culture, which is characterized by comprising the following steps:

(1) placing a cell culture glass slide in a glass slide culture box, wherein the side coated with polylysine faces upwards;

(2) fixing the separation frame on a cell culture glass slide, and performing separation culture on different cells in different holes of the separation frame; with the existence of polylysine, cells are relatively easy to adhere to the wall;

(3) after the cells adhere to the wall, separating the separating frame from the cell culture glass slide by using a separator, removing the separating frame, and still placing the glass slide in a glass slide culture box for culture;

(4) after the different cells removed from the spacer meet and contact with each other as the culture time increases, the cells interact with each other.

The technical scheme of the invention has the following beneficial effects: the slide glass system for direct contact cell co-culture has the advantages of simple structure, convenient use, simple operation, less required culture solution, economy and environmental protection.

Different cells are cultured in a separation mode through the separation frame on the cell culture glass slide, the cells are easy to adhere to the wall due to the existence of polylysine, after the cells adhere to the wall, the hole frame and the glass slide are separated by using the separator, the separation frame is removed, the cell culture glass slide is still placed in the glass slide culture box for culture, and compared with the conventional culture dish and the like for culture, the glass slide system for direct contact cell co-culture needs less culture solution, is environment-friendly, and saves resources.

In the invention, a four-hole glass slide system, a six-hole glass slide system or an eight-hole glass slide system can be selected according to different cells of experiment targets, and inner baffles (0.1-0.5 mm) with different thicknesses can be selected according to different lengths according to research requirements.

In the invention, when different cells with the separation frame removed meet and contact with each other along with the prolonging of the culture time, the interaction between the cells begins to be generated, and the detection of the cells can be carried out by simple observation or immunofluorescence staining of various proteins, and the research of different treatments (such as silencing or overexpression) can be carried out on the basis, thereby completely meeting the requirements of both contact co-culture and subsequent research.

Drawings

FIG. 1 is a schematic structural view of a four-well slide system according to example 1 of the present invention;

FIG. 2 is a schematic structural view of a six-well slide system in example 2 of the present invention;

FIG. 3 is a schematic structural view of an eight-well slide system in example 2 of the present invention;

FIG. 4 is a schematic view of the slide culture cassette of the present invention;

FIG. 5 is a schematic view of the separator of the present invention.

Description of reference numerals: 1. cell culture slides; 11. sanding the single plate; 2. a spacer; 21. an outer frame; 22. an inner partition plate; 3. a slide culture box; 31. a box body; 32. a cover body; 4. a separator.

Detailed Description

In order to make the technical problems, technical solutions and advantages of the present invention more apparent, the following detailed description is given with reference to the accompanying drawings and specific embodiments.

In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", "front", "rear", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of description and simplicity of description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.

In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.

A slide glass system for direct contact cell co-culture comprises a slide glass culture box 3, a cell culture slide glass 1 arranged on the slide glass culture box 3, and a separation frame 2 which is detachably arranged on the cell culture slide glass 1. The invention also has a separator 4 for separating the spacer 2 from the cell culture slide 1.

In a further embodiment, the slide culture box 3 comprises a box body 31 and a cover 32 which is detachably arranged above the box body 31, wherein the cover 32 is matched with the box body 31 in size; the slide system also includes a separator 4 for separating the spacer 2 from the cell culture slides 1. In the invention, the slide glass culture box 3 is beneficial to taking, placing and moving at the super clean bench, the incubator, the microscope table surface and the like, and the cover body 32 made of the same material is arranged on the box body 31 of the slide glass culture box 3, thus playing the roles of preventing cell pollution and reducing the evaporation of culture medium.

Preferably, the left side or the right side of the cell culture slide glass 1 is provided with a single-side frosted surface 11, and the upper end surface of the cell culture slide glass 1 is coated with polylysine. In the invention, one side of the cell culture glass slide 1 is provided with the single-side frosted surface 11, so that the situation of mixing left and right sample adding is avoided, and the cell culture glass slide 1 not only can culture adherent cells, but also can be directly used for observation of an inverted microscope or an inverted laser confocal microscope.

In a further embodiment, the spacer 2 is provided with a biocompatible acrylic adhesive on its lower end face, and the spacer 2 is sealingly connected to the cell culture slide 1 by an adhesive substance.

In a further embodiment, the cell culture slide 1 is made of white borosilicate glass material, and the cell culture slide 1 has a size of 25mm × 75mm and a thickness of 1-1.2 mm.

In a further embodiment, the spacer 2 is made of plastic material.

In a further embodiment, the partition frame 2 is divided into an outer frame 21 and an inner partition 22, the thickness of the outer frame 21 is 1mm, the thickness of the inner partition 22 is 0.1-0.5mm, and preferably, the thickness of the inner partition is 0.1mm, 0.3mm or 0.5 mm; the height of the outer frame 21 and the inner partition 22 is 10 mm.

In a further embodiment, the partition shelf 2 may be divided into three types, i.e., a four-hole shelf, a six-hole shelf and an eight-hole shelf according to the number of holes, wherein the sizes of the four holes in the four-hole shelf are all 12mm × 23mm, the sizes of the six holes in the six-hole shelf are all 8mm × 23mm, and the sizes of the eight holes in the eight-hole shelf are all 12mm × 11.35 mm.

In a further embodiment, the slide culture box 3 is made of transparent polystyrene, and the dimensions of the slide culture box 3 are 83mm × 30mm × 15 mm.

The embodiment of the invention also provides a working method of the slide system for the direct contact cell co-culture, which comprises the following steps:

(1) placing a cell culture slide glass 1 in a slide glass culture box 3, wherein the side coated with polylysine faces upwards;

(2) fixing the separation frame 2 on the cell culture glass slide 1, and performing separation culture on different cells in different holes of the separation frame 2; with the existence of polylysine, cells are relatively easy to adhere to the wall;

(3) after the cells adhere to the wall, the separating frame is separated from the cell culture glass slide by using a separator 4, the separating frame is removed, and the glass slide is still placed in a glass slide culture box for culture;

(4) after the different cells removed from the spacer 2 meet and contact with each other as the culture time increases, the cells interact with each other.

Example 1

As shown in FIG. 1, a direct contact cell co-culture slide system comprises a polylysine coated cell culture slide 1 and a four-well spacer 2, collectively referred to as a four-well slide system, typically housed in a slide culture cassette 3 as shown in FIG. 4. When the interaction between two kinds of cells is researched, different kinds of cells are planted in adjacent holes of the four-hole separation frame 2 at intervals, the cover bodies 32 of the corresponding glass slide culture boxes 3 can be marked, and the added cell sequence can be remembered through the single-side frosted surface 11, so that confusion is avoided. After the cells adhere to the wall, the separator 4 shown in fig. 5 is used for separating the separating frame 2 from the cell culture glass slide 1, the separating frame 2 is removed, the cell culture glass slide 1 is still placed in the glass slide culture box 3 for culture, the two cells can be directly contacted after a period of time, a special structure is formed or other reactions are initiated after interaction or treatment, and the junction of the two cells can be directly observed or correspondingly detected and analyzed.

Example 2

As shown in fig. 2 and 3, a slide system for direct contact cell co-culture includes a cell culture slide 1 coated with polylysine and a six-well partition rack 2 or an eight-well partition rack 2, which are referred to as a six-well slide system and an eight-well slide system, respectively. Different specifications of slide systems can be selected according to different cells of experiment purposes. When the growth of cells is relatively slow, a six-well slide system or even an eight-well slide system can be selected; when the experiment needs to be performed once and the verification is repeated, the two glass slide systems can be selected, and the experiment can be repeated at least for more than three times; when the interaction of one cell and two cells needs to be studied simultaneously, an eight-hole glass slide system can be selected and planted in the left and right and upper or lower adjacent holes respectively; when the cells under study will protrude, the inner baffles (0.1-0.5 mm) can be chosen to have different thicknesses depending on the length.

While the foregoing is directed to the preferred embodiment of the present invention, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (10)

1. A slide glass system for direct contact cell co-culture is characterized by comprising a slide glass culture box, a cell culture slide glass arranged on the slide glass culture box, and a separation frame which is detachably arranged on the cell culture slide glass; the slide system also includes a separator for separating the spacer from the cell culture slides.

2. A direct contact cell co-culture slide system as claimed in claim 1, wherein the slide culture cassette includes a cassette body and a cover detachably disposed over the cassette body, the cover being sized to fit the cassette body.

3. A direct contact cell co-culture slide system as claimed in claim 1, wherein one side of the cell culture slide is provided with a single frosted surface, and the upper surface of the cell culture slide is coated with polylysine.

4. A direct contact cell co-culture slide system as claimed in claim 1, wherein the spacer is provided with a biocompatible acrylic adhesive on its lower end surface, and the spacer is sealingly connected to the cell culture slide by the adhesive.

5. A direct contact cell co-culture slide system as claimed in claim 1, wherein said cell culture slide is made of white borosilicate glass, said cell culture slide having dimensions of 25mm x 75mm and a thickness of 1-1.2 mm.

6. A direct contact cell co-culture slide system as claimed in claim 1, wherein the spacer is made of transparent polystyrene.

7. The slide system for direct contact cell co-culture according to claim 1, wherein the separation frame is divided into an outer frame and an inner partition, the outer frame has a thickness of 1mm, the inner partition has a thickness of 0.1mm, 0.3mm or 0.5mm, and the height of the outer frame and the inner partition is 10 mm.

8. A direct contact cell co-culture slide system as claimed in claim 1, wherein the partition shelf is divided into three kinds of four-well shelf, six-well shelf and eight-well shelf, the size of the hole in the four-well shelf is 12mm x 23mm, the size of the hole in the six-well shelf is 8mm x 23mm, and the size of the hole in the eight-well shelf is 12mm x 11.35 mm.

9. A direct contact cell co-culture slide system as claimed in claim 1, wherein the slide culture box is made of transparent polystyrene material, and the size of the slide culture box is 83mm x 30mm x 15 mm.

10. A method of operating a slide system for direct contact cell co-culture according to claim 1, comprising the steps of:

(1) placing a cell culture glass slide in a glass slide culture box, wherein the side coated with polylysine faces upwards;

(2) fixing the separation frame on a cell culture glass slide, and performing separation culture on different cells in different holes of the separation frame; with the existence of polylysine, cells are relatively easy to adhere to the wall;

(3) after the cells adhere to the wall, separating the separating frame from the cell culture glass slide by using a separator, removing the separating frame, and still placing the glass slide in a glass slide culture box for culture;

(4) after the different cells removed from the spacer meet and contact with each other as the culture time increases, the cells interact with each other.

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