CN210596107U

CN210596107U - Adherent unicellular double-deck culture apparatus that divides

Info

Publication number: CN210596107U
Application number: CN201921573677.9U
Authority: CN
Inventors: 李中原; 郭海婷; 谭洁; 李鸿文; 甄瑞峰; 夏彬; 李夏
Original assignee: Guilin Medical University
Current assignee: Guilin Medical University
Priority date: 2019-09-20
Filing date: 2019-09-20
Publication date: 2020-05-22
Anticipated expiration: 2029-09-20

Abstract

The utility model discloses an adherence unicell divides double-deck culture apparatus, include slide glass, inserted sheet, plectrum, drive strip, carrier frame and gu the post, it is located the device top to drive the strip, and the below is the plectrum, the inserted sheet includes inserted sheet flank, top cap and hinge region, and there is the support that prevents the adhesion of two wings bottom, and the inserted sheet flank links to each other with the top cap through the hinge region, the slide glass is the cavity slide, including carrying groove, edge hole, carrying groove support, slide glass inside and outside frame and last lower supporting arris, gu the long square post of post for fixed slide glass, the carrier frame is the three-dimensional frame including edge post and solid post draw-in groove. The utility model discloses adherence unicell divides double-deck complicated device of cultivateing, application earth gravity and adherence cell adherence growth characteristic, the bicell that divides the gained for the first time adherence unicell of will adhering to the wall is on being the inserted sheet flank of mirror image through upright hatching and handstand cultivation. The device has the advantages of fine structure, ingenious design, simple and convenient operation, high double-layer culture efficiency and convenience for laboratory research and sterile operation.

Description

Adherent unicellular double-deck culture apparatus that divides

Technical Field

The utility model belongs to the technical field of general chemistry or physics laboratory paraphernalia, in particular to adherence unicellular double-deck culture apparatus that divides.

Background

Adherent cells (adherentcells) are a class of eukaryotic cell lines that rely on self-secretion, culture fluid supply, or adhesion factors coated on the surface of a support to grow and propagate on the surface of the support. The growth process of adherent cells is mainly divided into a free phase, an adherent phase, a latent phase, a logarithmic phase, a plateau phase and a decline phase, and after the adherence, epithelial cell-like or fibroblast-like growth forms are generally formed. Adherent cells have become important laboratory materials for researching vaccines, developing new drugs, preparing monoclonal antibodies, analyzing gene functions and drug action mechanisms, culturing pathogenic microorganisms in vitro and detecting disease occurrence and development mechanisms and the activity rules of adherent cells, and the scientific research value of the adherent cells is not light.

At present, the commonly used adherent cell culture method is as follows: after the cells to be passaged are digested by pancreatin, the cells are blown into single cell suspension, then the single cell suspension is paved into a cell bottle or a culture plate according to the specified inoculation proportion, and the cell suspension is placed into a cell culture box to continue the camp adherent growth until the experiment requirements are met or the bottom surface of a support is fully distributed. It is obvious that the adherent cells obtained by the above-mentioned passage method are actually cell groups obtained by multiple mitosis of adherent single cells with unknown number, and the cell spacing between the adherent single cells in each cell bottle or culture plate is not equal except for the difference between the initial cell laying number and the mitosis number. The obtained adherent cells are also a cell population which is uniform in morphology, stable in growth and clear in biological characters, and the propagation activity and the metabolic capability of the adherent cells are obviously different.

SUMMERY OF THE UTILITY MODEL

In order to solve the technical problem, the utility model provides an adherent single cell divides double-deck culture apparatus, including slide glass, inserted sheet, plectrum, drive strip, carrier frame and solid post, drive strip and lie in the apparatus top while being said, including strip head and strip body; the poking piece is positioned below the driving strip and comprises a piece body, a clamping part, a poking piece ring, a rotary column and a rotary column base, wherein the poking piece ring is a buckle for connecting the inserting piece, the poking piece can rotate relative to the inserting piece, and the piece body is a glass slide for poking the side wing of the inserting piece; the insert comprises an insert side wing, a top cover and a hinge region, wherein a support column for preventing double wings from being adhered is arranged at the bottom of the side wing, a pressing edge is positioned on the inner side above the side wing, the height of the pressing edge is the same as that of the support column, and the insert side wing is connected with the top cover through the hinge region; the slide glass is a hollow slide glass and comprises a slide groove, edge holes, a slide groove bracket, slide glass inner and outer frames and upper and lower support edges, wherein the edge holes are used for inserting edge columns, and the support edges are square columns for supporting the slide glass, assisting in fixing the inserting sheets and keeping the distance between the adjacent slide glasses; the fixed column is a rectangular column for fixing the slide glass; the carrier frame is a three-dimensional frame comprising edge columns and column fixing clamping grooves.

Preferably, the driving strips are double strips with opposite movement directions, the strip heads are pinching parts, anti-slip grains are arranged on the surfaces of the strip heads, the strip bodies are provided with circular strip holes, and the number of the strip bodies is the same as that of the driven poking pieces.

Preferably, the rotating column of the shifting piece is a round column inserted into the strip hole, the height of the rotating column is the same as the thickness of the driving strip, and the diameter of the rotating column is smaller than that of the rotating base; the external diameter of the rotating column is slightly smaller than the internal diameter of the strip hole, and the driving strip and the shifting piece can rotate around the rotating column.

Preferably, the clamping part of the shifting piece is positioned in the middle of the upper part of the shifting piece, the height of the clamping part is the same as that of the rotating base, and the clamping part is used for clamping and lifting by hands.

Preferably, the middle part of the top cover is provided with two shifting piece jacks, and four corners of the top cover are respectively provided with a circular top column.

Preferably, the side wings of the inserting piece are left and right double wings in mirror image, and the inner side surfaces of the double wings are provided with adherent cell adhesion zones.

Preferably, the carrier is a U-shaped channel capable of carrying only a single adherent cell.

The utility model provides a divide double-deck new installation of cultivateing adherence unicell to and a adherence unicell divides double-deck new feasibility scheme of cultivateing.

The utility model is designed according to the earth gravity and the adherent growth characteristics of adherent cells, firstly, the carrier frame with the slide glass is immersed into the adherent cell suspension, and is taken out after acting for 3-5 s; inserting the inserting sheet and the shifting sheet, installing the driving strip and the fixing column, and after the cells are completely attached to the wall, inverting the culture device until the first mitosis is completed; taking out the solid column after the connection between the pancreatin digestive double cells, pulling out the insert in the positive direction, and developing and culturing. The utility model discloses the structure is meticulous, design benefit, and the adherence unicellular divides double-deck efficient, the laboratory research and the aseptic technique of being convenient for.

Drawings

The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.

FIG. 1 is a perspective view of an adherent single cell double-layer culture device according to an embodiment of the present invention

FIG. 2 is a top view of the adherent single cell double-layer culture apparatus according to the embodiment of the present invention.

FIG. 3 is a perspective view of the adherent single cell double-layered culture apparatus according to the embodiment of the present invention.

FIG. 4 is a schematic cross-sectional view of an adherent single cell double-layer culture apparatus according to an embodiment of the present invention.

Figure 5 is a top view of a slide.

Figure 6 is a longitudinal and cross-sectional schematic of a slide.

FIG. 7 is a schematic top view of the tabs erected, flared and flared.

Fig. 8 is a perspective view of the paddle.

Fig. 9 is a schematic longitudinal section of the pick.

Fig. 10 is a schematic structural diagram of the driver blade.

Fig. 11 is a perspective view of the fixing post.

Fig. 12 is a perspective view of the carriage.

FIG. 13 is a schematic view of the operation process of the adherent single cell double-layer culture device according to the embodiment of the present invention.

In the figure, 1: carrying a slide; 2: inserting sheets; 3: a shifting sheet; 4: driving the strips; 5: a carrier; 6: fixing the column; 11: a carrying groove; 12: a carrier trough bracket; 13: a slide inner frame; 14: upper supporting edges; 15: lower supporting edges; 16: a slide outer frame; 17: an edge hole; 18: a cell-bearing region; 21: inserting sheet side wings; 22: supporting columns; 23: pressing edges; 24: a top cover; 25: a top pillar; 26: a plectrum jack; 27: an adhesion zone; 28: a hinge region; 31: a sheet body; 32: a clamping portion; 33: a disc pulling ring; 34: a spin column; 35: a column base; 41: strip ends; 42: a strip body; 43: a bar hole; 44: anti-skid lines; 51: an edge post; 52: a clamping groove for fixing the column.

Detailed Description

As shown in figures 1-13, the utility model discloses an adherent single cell divides double-deck culture device, is a slide combination type container for dividing double-deck culture adherent single cell.

The adherence is the characteristic that the adherence can grow and propagate on the surface of the support only by means of the secretion of cells per se, the supply of culture solution or the coating of adhesion factors on the surface of a cell support.

The single cell is a single cell group obtained after the adherent cell to be transmitted is digested by pancreatin.

The double-layer culture is a culture mode of separating double cells obtained by first mitosis of single cells in a mirror image mode by using the characteristics of earth gravity and adherent cell adherent growth.

The container is a vitreous container.

An adherent single cell divides double-deck culture apparatus, including slide glass 1, male tab 2, plectrum 3, drive strip 4, carrier 5 and solid post 6, drive strip 4 lie in apparatus top while being said, including strip head 41 and strip body 42; the plectrum 3 is positioned below the driving strip 4 and comprises a sheet body 31, a clamping part 32, a plectrum ring 33, a rotating column 34 and a rotating column base 35, the plectrum ring 33 is a buckle for connecting the inserting sheet 2, the plectrum can rotate relative to the inserting sheet, and the sheet body 31 is a glass sheet for shifting the inserting sheet flank 21; the insert 2 comprises an insert side wing 21, a top cover 24 and a hinge area 28, a support pillar 22 for preventing double wings from adhesion is arranged at the bottom of the side wing, a pressing edge 23 is positioned on the inner side above the side wing, the height of the pressing edge is the same as that of the support pillar 25, the insert side wing 21 is connected with the top cover 24 through the hinge area 28, and the hinge can be made of a flexible material capable of being folded back; the lower part of the sheet body is positioned below the top cover 24 and is used for shifting the inserting sheet side wings 21; the slide glass 1 is a hollow slide glass and comprises a slide groove 11, an edge hole 17, a slide groove bracket 12, slide inner and

outer frames

13 and 16, and upper and

lower support ribs

14 and 15, wherein the edge hole 17 is used for inserting an edge column 51, and the support ribs are square columns for supporting the slide glass 1, assisting in fixing the insert 2 and keeping the distance between the adjacent slide glasses 1; the fixed column 6 is a rectangular column for fixing the slide glass 1; the carrier 5 is a three-dimensional frame comprising an edge column 51 and a column fixing clamping groove 52.

The driving strips 4 are double-piece strips with opposite movement directions, the strip heads 41 are holding parts, anti-skid grains 44 are arranged on the surfaces, and the strip bodies 42 are provided with circular strip holes 43, and the quantity of the driving strips is the same as that of the driven poking pieces 3.

The rotating column 34 of the shifting piece 3 is a round column inserted into the strip hole 43, the height of the rotating column is the same as the thickness of the driving strip 4, and the diameter of the rotating column is smaller than that of the rotating base; the outer diameter of the rotating column 34 is slightly smaller than the inner diameter of the strip hole 43, and the driving strip and the shifting piece can rotate around the rotating column.

The clamping part 32 of the plectrum 3 is positioned in the middle of the upper part of the plectrum 3, the height of the clamping part 32 is the same as that of the rotary base, and the clamping part 32 is used for clamping and lifting by hands.

The top cover 24 has two shifting piece jacks 26 in the middle, and a circular top column 25 is arranged at each of the four corners.

The side wings 21 of the inserting sheet are left and right double wings in mirror image, and the inner side surfaces of the double wings are provided with adherent cell adhesion zones 27.

The carrier groove 11 is a U-shaped channel groove which can only bear single adherent cells.

The operation flow is as follows:

1. preparing adherent single cell suspension;

1) removing the upper culture solution when adherent cells are fully distributed on the bottom surface of the support, adding appropriate amount of 0.25% pancreatin solution, and standing CO₂Digesting in an incubator until the cells completely fall off.

2) Adding appropriate amount of cell culture solution (containing 10% fetal calf serum and 1 Xdouble antibody), mixing, blowing to obtain single cell state, and transferring into sterile 50mL centrifuge tube, 6-well or 12-well culture plate.

2. The utility model is assembled and disassembled and cells are cultured;

1) the slides 1 are loaded in succession along the edge column 51 into the carrier 5, the carrier with the slides is immersed upright in the above-mentioned single-cell suspension and taken out after 3-5s of action.

2) The double wings of the inserting piece are carefully inserted into the side grooves of the lower supporting edges (the area between the lower supporting edges and the inner frame 13 of the slide glass), the shifting piece 3 is installed (the direction of the piece body 31 is parallel to the side wings of the inserting piece 2), and then the inserting piece is pushed downwards to the bottom of the carrier frame 5.

3) The insert 2 is then pushed forward as indicated in figure 13 to the upper support ridge 14, and the two wings of the insert 2 are gripped by sterile forceps and held tightly against the intermediate loading slot 11.

4) The insert 2 is pushed forward into the side groove of the upper supporting rib 14 (the area between the upper supporting rib 14 and the carrying groove bracket 12), the fixed column 6 is inserted into the fixed column clamping groove 52, and then the incubation culture is continued in a proper amount of cell culture solution (containing 10% fetal calf serum and 1 multiplied double antibody).

5) After the cells are fully attached to the wall, the device is inverted and cultured until the first mitosis of the cells is completed.

6) After the connection between the two cells obtained by digestion with a proper amount of pancreatin, the fixing column 6 is taken out, the inserting piece 2 is pulled backwards to the lower support edge, and the driving strip 4 is installed and pulled towards the two sides.

7) When the side wings of the inserting pieces 2 abut against the slide inner frame 13, the driving strips 4 are detached, and the inserting pieces 2 are pulled backwards into the lower support edge side grooves.

8) Taking out the shifting sheet 3, pulling out the inserting sheet 2 upwards, placing the inserting sheet 2 in a cell bottle or a culture plate, and performing culture in a mode that the cell adhesion zone 27 is upwards.

9) According to the research requirement, if a large number of adherent cells in mirror image are needed, the size of the side wing of the insert can be properly increased and/or the number of the slide glass 1 and the insert glass 2 can be increased.

Adherent cells are a eukaryotic cell line which can grow and propagate on the surface of a support only by virtue of an adhesion factor which is secreted by the cells and provided by a culture solution or coated on the surface of the support, and become important laboratory materials for researching vaccines, developing new drugs, preparing monoclonal antibodies, analyzing gene functions and drug action mechanisms, culturing pathogenic microorganisms in vitro and detecting disease occurrence and development mechanisms and the self-activity rules of the adherent cells. The obtained adherent cells are actually cell groups obtained by multiple mitosis of adherent unicells with unknown numbers by applying a traditional passage mode, and the cell spacing between the adherent unicells in each cell bottle or culture plate is unequal except that the initial cell laying number and the mitosis times are different, so that the reproductive activity and the metabolic capacity of the obtained adherent cells are obviously different. The utility model provides a complicated device that adherence unicell divides double-deck cultivation utilizes earth gravity and adherence cell adherence growth characteristic, and the bicell that will adhere to the first division of adherence unicell gained is adhered on being the inserted sheet flank of mirror image through upright hatching and handstand cultivation. The device has the advantages of fine structure, ingenious design, simple and convenient operation, double-layer division, high efficiency and convenience for laboratory research and aseptic operation.

Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or equivalents may be substituted for elements thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The utility model provides an adherence unicell divides double-deck culture apparatus which characterized in that: the device comprises a slide (1), an inserting sheet (2), a shifting sheet (3), a driving strip (4), a carrier (5) and a fixing column (6), wherein the driving strip (4) is positioned above the device and comprises a strip head (41) and a strip body (42); the poking piece (3) is positioned below the driving strip (4) and comprises a piece body (31), a clamping part (32), a poking piece ring (33), a rotating column (34) and a rotating column base (35), the poking piece ring (33) is a buckle connected with the inserting piece (2), the poking piece can rotate relative to the inserting piece, and the piece body (31) is a glass slide for poking the inserting piece side wing (21); the inserting piece (2) comprises an inserting piece side wing (21), a top cover (24) and a hinge area (28), a supporting column (22) for preventing double wings from being adhered is arranged at the bottom of the side wing, the pressing edge (23) is positioned on the inner side above the side wing, and the inserting piece side wing (21) is connected with the top cover (24) through the hinge area (28); the slide glass (1) is a hollow slide glass and comprises a slide groove (11), edge holes (17), a slide groove bracket (12), inner and outer frames (13, 16) of the slide glass, and upper and lower support edges (14, 15), wherein the edge holes (17) are used for inserting edge columns (51), and the support edges are square columns for supporting the slide glass (1), assisting in fixing the inserting sheets (2) and keeping the distance between the adjacent slide glasses (1); the fixed column (6) is a rectangular column for fixing the slide glass (1); the carrier (5) is a three-dimensional frame comprising edge columns (51) and column fixing clamping grooves (52).

2. The adherent single cell bilayer culture device of claim 1, wherein: the driving strips (4) are double strips with opposite movement directions, the strip head (41) is a pinching part, the surface of the strip head is provided with anti-slip grains (44), the strip body (42) is provided with circular strip holes (43), and the quantity of the strip heads is the same as that of the driven plectrums (3).

3. The adherent single cell bilayer culture device of claim 1, wherein: the rotating column (34) of the shifting piece (3) is a round column inserted into the strip hole (43), the height of the rotating column is the same as the thickness of the driving strip (4), and the diameter of the rotating column is smaller than that of the rotating base; the outer diameter of the rotating column (34) is slightly smaller than the inner diameter of the strip hole (43), and the driving strip (4) and the shifting piece (3) can rotate around the rotating column.

4. The adherent single cell bilayer culture device of claim 1, wherein: the clamping part (32) of the shifting piece (3) is positioned in the middle of the upper part of the shifting piece (3) and is as high as the rotary base, and the clamping part (32) is used for clamping and lifting by hands.

5. The adherent single cell bilayer culture device of claim 1, wherein: two shifting piece insertion holes (26) are formed in the middle of the top cover (24), and a circular top column (25) is arranged at each of four corners of the top cover.

6. The adherent single cell bilayer culture device of claim 1, wherein: the inserting sheet side wings (21) are mirror-image left and right double wings, and the inner side surfaces of the double wings are provided with adherent cell adhesion zones (27).

7. The adherent single cell bilayer culture device of claim 1, wherein: the carrier groove (11) is a U-shaped channel groove which can only carry single adherent cells.