CN212476779U - Multi-channel 3D cell invasion transfer measuring device - Google Patents

Abstract

The multi-channel 3D cell invasion transfer determination device comprises a matrigel injection cylinder, a cell invasion transfer box, four cell culture boxes and four slide storage boxes, wherein the cell invasion transfer box and the four cell culture boxes are of box body structures in cuboid appearances, and the tops of the cell invasion transfer box and the four cell culture boxes are open; in summary, the utility model injects matrigel by using the matrigel injection cylinder, and the four matrigel solution cavities of the matrigel injection cylinder are mutually independent and not communicated with each other, so as to prevent the cross influence of different cells or cell factors; the utility model discloses convenient to use, cell or different cytokines are to the influence that the invasion and attack was shifted in can a plurality of different cell culture liquids of simultaneous determination, and the convenience is drawn materials to matrigel inner cell, and convenient regularly observation prevents the interact.

Description

Multi-channel 3D cell invasion transfer measuring device

Technical Field

The utility model relates to a multichannel 3D cell invasion and attack transfer survey device.

Background

Tumor invasion is the process of infiltration of tumor cells from their site of origin along the interstitial space into the surrounding tissue, and is marked by the breakthrough of tumor cells through the basement membrane. Tumor invasion is the result of tumor cell, peripheral stromal interactions, and is the first step in tumor spread. Tumor metastasis is the process by which malignant tumor cells invade lymphatic vessels, blood vessels, or body cavities from the primary site, migrate to a new location, and generate new or secondary tumors in the new environment. Unlike the planar growth mode (2D) of cell lines, 3D cells are grown in a three-dimensional form, including cell microspheres (sphenoid) and organoids (organoids). The 3D cell is closer to the cell growth mode of human cells, so that the in-vivo state of the tumor cell can be reflected better.

To study the mechanism of tumor cell invasion and metastasis and to develop drugs that inhibit invasion and metastasis, we have used in vitro cell culture to mimic the environment of a living organism. 2D cells grow on a plane, which is a great difference from the way in which living organisms grow. 3D cell culture is a novel cell culture method, and 3D cell culture models can more accurately represent the natural environment experienced by cells in living organisms. Therefore, simulating the invasion and metastasis of tumor cells in vitro has important significance.

Invasive metastasis is an important feature of tumor cells and is also a major cause of death in patients due to tumors. The 3D cell invasion and metastasis experiment is mainly used for simulating the biological behavior of tumor cell invasion and metastasis in vitro. The 3D cell invasion and metastasis experiment plays an important role in researching the development mechanism of tumors and screening tumor drugs. Firstly, a 3D cell invasion and metastasis experiment can be used for researching the interaction between the tumor and surrounding interstitium (fibroblasts, immune cells and the like); secondly, 3D cell invasion and metastasis can be used for researching a specific mechanism in the process of generating and developing tumor cells; thirdly, 3D cell invasion and metastasis can be used for researching and developing drugs for inhibiting invasion and metastasis, in vitro experiments are important steps in drug research and development, and tumor drug research can be greatly promoted.

Currently, 2D cultured cells use a Transwell method to detect invasion and metastasis of the cells; the method for detecting Invasion and metastasis of 3D cultured cells is few, the detection method of Trevigen company (Cultrex 3D spherical Cell Invasion Assay) does not have a special detection kit, but extracellular matrix (equivalent to matrigel) is directly added into a 96-well plate, then fluorescence is added into a culture solution to mark cells, the morphological change of the cells is observed through a special instrument, and finally, detection software is needed to measure and calculate the result; the detection method is simple in design, the cells need to be marked by adding fluorescence, the cells are possibly damaged by adding the fluorescence, the experimental result is influenced, and the method needs a special instrument to track the cells marked by the fluorescence, so that the detection cost is improved; in addition, this method is currently unable to detect cell-to-cell interactions.

SUMMERY OF THE UTILITY MODEL

The utility model discloses a solve the weak point among the prior art, provide a multichannel 3D cell invasion and attack transfer survey device that can the multichannel study cell invasion and attack transfer, and each other do not influence.

In order to solve the technical problem, the utility model adopts the following technical scheme: a multi-channel 3D cell invasion transfer determination device comprises a matrix glue injection cylinder, a cell invasion transfer box, four cell culture boxes and four slide storage boxes, wherein the cell invasion transfer box and the four cell culture boxes are of box body structures in the shape of a cuboid, the tops of the cell invasion transfer box and the four cell culture boxes are open, the four cell culture boxes are fixedly connected to four side surfaces of the cell invasion transfer box respectively, the matrix glue injection cylinder comprises a partition plate part, an upper cylinder body and a lower cylinder body, the upper cylinder body is of a cylinder structure, the lower cylinder body is of a circular truncated cone structure with a large upper part and a small lower part, the upper cylinder body is fixed at the upper end of the lower cylinder body, the lower cylinder body is fixed at the center of the bottom of the cell invasion transfer box, the cross section of the partition plate part is cross-shaped, the partition plate part is fixed in the upper cylinder body and the lower cylinder body, the partition plate part divides an inner cavity, the four matrigel solution cavities correspond to the four cell culture boxes, the four cell culture boxes are respectively connected with the lower cylinder body of the matrigel injection cylinder through a transfer channel, and each cell culture box is communicated with the corresponding matrigel solution cavity through the transfer channel;

the slide storage box comprises an inserting rod piece, an outer stop block and an inner inserting box, wherein the outer stop block is fixed at the end part of the inner inserting box, the appearance of the inner inserting box is matched with the transfer channel, a through hole is formed in the outer stop block, an inserting hole is formed in the outer stop block along the vertical direction, the inner inserting box is of a box body structure with a through front part and a through back part, the inner cavity of the inner inserting box is correspondingly communicated with the through hole, three pairs of inserting groove groups are arranged on the box walls opposite to two sides of the inner inserting box, two slides are inserted into each group of inserting groove groups, a frosted handheld part is arranged at the end part of; the inserting rod piece comprises an operating rod and a plugging plate, the operating rod is fixed at the top of the plugging plate, the plugging plate plugs the through hole by inserting into the jack of the outer baffle, and the cross section of the plugging plate is of a wedge-shaped structure;

the wall of each cell culture box is provided with a marking plate and a timer.

And sealing rubber layers are arranged on the outer surfaces of the plugging plate and the inner inserting box.

Adopt above-mentioned technical scheme, the utility model discloses following beneficial effect has: the utility model uses the matrigel injection cylinder to inject the matrigel, and the four matrigel solution cavities of the matrigel injection cylinder are mutually independent and are not communicated with each other, so as to prevent the cross influence of different cell factors; the utility model discloses convenient to use, cytokine is to the influence that the invasion and attack was shifted in can a plurality of different cell culture liquid of simultaneous determination, and the convenience is gathered materials the matrigel inner cell, makes things convenient for regularly to observe, prevents the mutual influence.

Drawings

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

FIG. 2 is a schematic view of the structure of the slide storage cassette;

FIG. 3 is a cross-sectional view of the inner cartridge;

FIG. 4 is a schematic view of the structure of a slide.

Detailed Description

As shown in FIGS. 1-4, the multi-channel 3D cell invasion transfer measuring device of the present invention comprises a matrigel injection cylinder 1, a cell invasion transfer box 2, four cell culture boxes 3 and four slide storage boxes, wherein the cell invasion transfer box 2 and the four cell culture boxes 3 are both rectangular box structures, the tops of the cell invasion transfer box 2 and the four cell culture boxes 3 are open, the four cell culture boxes 3 are respectively and fixedly connected to four sides of the cell invasion transfer box 2, the matrigel injection cylinder 1 comprises a partition plate 4, an upper cylinder 5 and a lower cylinder 6, the upper cylinder 5 is a cylinder structure, the lower cylinder 6 is a circular truncated cone structure with a large upper part and a small lower part, the upper cylinder 5 is fixed at the upper end of the lower cylinder 6, the lower cylinder 6 is fixed at the bottom center of the cell invasion transfer box 2, the cross section of the partition plate 4 is cross-shaped, the partition plate 4 is fixed in the upper cylinder 5 and the lower cylinder 6, the partition plate part 4 divides the inner cavity of the matrigel injection cylinder 1 into four matrigel solution cavities 7 which are independent from each other, the four matrigel solution cavities 7 correspond to the four cell culture boxes 3, the four cell culture boxes 3 are respectively connected with the lower cylinder body 6 of the matrigel injection cylinder 1 through a transfer passage 8, and each cell culture box 3 is communicated with the corresponding matrigel solution cavity 7 through the transfer passage 8;

the slide storage box comprises an inserting rod piece, an outer stop block 9 and an inserting box 10, wherein the outer stop block 9 is fixed at the end part of the inserting box 10, the appearance of the inserting box 10 is matched with the transfer channel 8, a through hole 11 is formed in the outer stop block 9, an inserting hole 12 is formed in the outer stop block 9 along the vertical direction, the inserting box 10 is of a box body structure which is transparent in the front and back direction, the inner cavity of the inserting box 10 is correspondingly communicated with the through hole 11, three pairs of inserting groove groups 13 are respectively arranged on the opposite box walls at two sides of the inserting box 10, two slides 14 are inserted into each group of inserting groove groups 13, a frosted handheld part 15 is arranged at the end part of each slide 14, and a; the rod inserting piece comprises an operating rod 17 and a plugging plate 18, the operating rod is fixed at the top of the plugging plate 18, the plugging plate 18 plugs the through hole 11 by inserting into the jack 12 of the outer baffle, and the section of the plugging plate 18 is of a wedge-shaped structure;

the wall of each cell culture chamber 3 is provided with a marking plate 19 and a timer 20.

The outer surfaces of the plugging plate 18 and the inner plug box 10 are provided with a sealing rubber layer.

The utility model is provided with a marking plate 19 and a timer 20 on each cell culture box 3, the marking plate 19 is used for marking the names, the concentrations and the like of different cell culture solutions, and the timer 20 can time the invasion and transfer degree of cells in different periods; when the cell invasion and transfer is carried out specifically, all the glass slides 14 are inserted into the slots one by one, wherein two glass slides 14 are in a group, one group of glass slides 14 are correspondingly inserted into the slot group 13 of the glass slide storage box, the end part of each glass slide 14 is provided with the frosted handheld part 15, the operation on the glass slides 14 is convenient, and the length scale marks 16 are arranged on the glass slides 14, so that the cell invasion and transfer distance is convenient to observe; after the glass slide 14 is assembled, the plugging plate 18 is inserted into the jack 12 of the outer baffle plate by holding the operating rod 17 by hand, the through hole 11 is sealed, then the glass slide storage box is placed into the cell culture box 3 by holding the operating rod 17 by hand, and the glass slide storage box is inserted into the transfer channel 8 connected with the matrigel injection cylinder 1 of the cell culture box 3;

then, matrigel in a dissolved state is poured into the four matrigel solution cavities 7 of the matrigel injection cylinder 1, the matrigel solution flows into the inner insertion box 10 of the transfer channel 8, at the moment, the inner insertion box 10 is completely filled with the matrigel solution, a sealing rubber layer on the outer surface of the inner insertion box 10 is used for sealing a gap between the inner insertion box 10 and the transfer channel 8, and a sealing rubber layer on the outer surface of the plugging plate 18 is used for sealing a gap between the plugging plate 18 and the hole wall of the insertion hole 12, so that the matrigel solution is prevented from entering the cell culture box 3 through the gap;

then the device is placed at the cell culture temperature, matrigel is solidified, then the insertion rod part is pulled out upwards, the through hole 11 is opened, culture solution containing different cells is poured into four cell culture boxes 3, the cells are cultured in the cell culture boxes 3 for a certain time, the cells in the matrigel are invaded and transferred under the action of different cytokines, after the experimental conditions are met, the cell culture solution is poured out, then the operation rod 17 is held by hands, the plugging plate 18 is inserted into the matrigel solution cavity 7, the matrigel close to one side of the transfer channel 8 is cut off by using the wedge-shaped structure of the plugging plate 18, then the plugging plate 18 is inserted into the outer baffle insertion hole 12, the insertion box 10 of the slide storage box is pulled out of the transfer channel 8 outwards, then the cell culture boxes 3 are pulled out upwards, at the moment, redundant matrigel possibly exists outside the insertion box 10, and the redundant matrigel outside the insertion box 10 is removed by using the plugging plate 18; the slides 14 are then pulled outward, with matrigel of the cell invasion metastasis present between the slides 14 and a set of slides 14, and the set of slides 14 is placed under a microscope to observe the condition in which the cell invasion metastasis.

In addition, Matrigel (Matrigel) can be dissolved at 4 ℃ and is in a liquid state, the Matrigel can be gradually solidified along with the rise of temperature, the Matrigel can be quickly gelatinized (about 10 minutes) generally at the temperature of 22-35 ℃, and can be gelatinized along with the reduction of temperature, so the Matrigel can be stored in a refrigerator at-20 ℃; when in dissolution, the gel is thawed and dissolved at 4 ℃, the matrigel can be operated by the liquid transfer tube, the suction head and the small tube after thawing and dissolution, and the matrigel after gelling can be in a liquid state again after 24 to 48 hours at 4 ℃.

The present embodiment is not intended to limit the shape, material, structure, etc. of the present invention in any form, and all of the technical matters of the present invention belong to the protection scope of the present invention to any simple modification, equivalent change and modification made by the above embodiments.

Claims (2)

1. Multichannel 3D cell invasion transfer assay device which characterized in that: the cell invasion and transfer box comprises a matrigel injection cylinder, a cell invasion and transfer box, four cell culture boxes and four slide storage boxes, wherein the cell invasion and transfer box and the four cell culture boxes are of box structures with cuboid appearances, the tops of the cell invasion and transfer box and the four cell culture boxes are open, the four cell culture boxes are respectively and fixedly connected with four side surfaces of the cell invasion and transfer box, the matrigel injection cylinder comprises a partition plate part, an upper cylinder body and a lower cylinder body, the upper cylinder body is of a cylindrical structure, the lower cylinder body is of a round platform structure with a large upper part and a small lower part, the upper cylinder body is fixed at the upper end of the lower cylinder body, the lower cylinder body is fixed at the center of the bottom of the cell invasion and transfer box, the cross section of the partition plate part is cross-shaped, the partition plate part is fixed in the upper cylinder body and the lower cylinder body, the partition plate part divides an inner cavity of the matrigel injection cylinder, the four cell culture boxes are respectively connected with the lower cylinder body of the matrigel injection cylinder through a transfer channel, and each cell culture box is communicated with the corresponding matrigel solution cavity through the transfer channel;

the slide storage box comprises an inserting rod piece, an outer stop block and an inner inserting box, wherein the outer stop block is fixed at the end part of the inner inserting box, the appearance of the inner inserting box is matched with the transfer channel, a through hole is formed in the outer stop block, an inserting hole is formed in the outer stop block along the vertical direction, the inner inserting box is of a box body structure with a through front part and a through back part, the inner cavity of the inner inserting box is correspondingly communicated with the through hole, three pairs of inserting groove groups are arranged on the box walls opposite to two sides of the inner inserting box, two slides are inserted into each group of inserting groove groups, a frosted handheld part is arranged at the end part of; the inserting rod piece comprises an operating rod and a plugging plate, the operating rod is fixed at the top of the plugging plate, the plugging plate plugs the through hole by inserting into the jack of the outer baffle, and the cross section of the plugging plate is of a wedge-shaped structure;

the wall of each cell culture box is provided with a marking plate and a timer.

2. The multi-channel 3D cell invasion transfer assay device according to claim 1, characterized in that: and sealing rubber layers are arranged on the outer surfaces of the plugging plate and the inner inserting box.

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