CN112391287A

CN112391287A - Dynamic culture device for applying stress/strain gradient field to cells by utilizing magnetic force

Info

Publication number: CN112391287A
Application number: CN201911313609.3A
Authority: CN
Inventors: 王鑫; 万芷翎; 张春秋
Original assignee: Tianjin University of Technology
Current assignee: Tianjin University of Technology
Priority date: 2019-12-18
Filing date: 2019-12-18
Publication date: 2021-02-23

Abstract

The invention provides a dynamic culture device for applying a stress/strain gradient field to cells by using magnetic force, which comprises the following components: the base and the upright post bracket support the culture cavity; a culture chamber as a culture place for the cells; the silica gel culture substrate comprises magnetic beads inside, is provided for a cell growth place and external force loading, and is connected with the culture cavity through threads to ensure better sealing property; energizing a solenoid to provide magnetic force for magnetic beads in the silica gel substrate so as to load tension on the substrate; a peristaltic pump to circulate the cell culture fluid through the culture device; a culture solution collecting cavity and a link in the intermediate circulation process of the cell culture solution; a culture medium transfer tube for transferring a cell culture medium between the respective sections. The invention has the beneficial effects that: a dynamic cell culture device with a simple external force loading form and safety is developed, and the culture and observation of cells in a specific mechanical environment are realized.

Description

Dynamic culture device for applying stress/strain gradient field to cells by utilizing magnetic force

Technical Field

The invention relates to the technical field of biomedical cell engineering in-vitro culture, in particular to a dynamic cell culture device loaded with an external force, which is used for in-vitro culture of cells and relevant biomechanical research.

Background

In order to overcome the difference caused by the difference between the stress condition of the cells in the static culture outside the human body and the stress condition of the cells in the human body, a dynamic culture environment similar to the internal condition of the human body needs to be provided for the cells. The traditional cell culture dish can not provide a liquid environment for cell flowing and external force loading. The invention provides a novel dynamic cell culture device, which can load external force on a cell culture substrate through magnetic force under the condition of non-contact with the cell culture substrate and can realize the circulating flow of a cell culture solution. The invention has compact structure and simple operation, and can realize the simulation of mechanical loading condition and liquid environment of cells in human body to a great extent.

Disclosure of Invention

The invention aims to realize a novel external force loading mode for cells, provide a liquid growth environment for cell flow, realize dynamic culture of the cells by electrifying a solenoid and magnetic beads, and improve the convenience and the innovation of the device.

The technical scheme of the invention is as follows:

a cell culture membrane tensile stress applying device comprises a device base 1, a vertical column 2 with holes, a lower electrified solenoid 3, a nut 4, a bolt 5, an upper electrified solenoid 6, a top cover 7, a non-porous vertical column 8, a culture cavity cover 9, a cell culture cavity 10, a culture substrate upper clamping cover 11, a sealing rubber ring 12, a silica gel cell culture substrate 13, a culture substrate lower clamping cover 14 and a positioning pin 15.

The nut 4, the bolt 5, the culture cavity cover 9, the cell culture cavity 10, the clamping cover 11 above the culture substrate, the sealing rubber ring 12, the silica gel cell culture substrate 13 and the clamping cover 14 below the culture substrate form a place for cell growth. Wherein the culture substrate upper clamping cover 11 and the culture substrate lower clamping cover 14 tightly clamp the silica gel cell culture substrate 13 at the center through bolts at the periphery, and are in threaded connection with the inner side of a round hole at the lower end of the cell culture cavity 10 through threads on the culture substrate upper clamping cover 11, thereby jointly forming a cell growth place. The cell culture cavity 10 is provided with through holes at two sides and can be connected with a conduit to form a flowing cell culture solution environment. The base 1 and the top cover 7 of the device are connected by the upright post 2 with holes and the upright post 8 without holes through screw threads to form a supporting part of the whole device, the cell culture cavity 10 is provided with through holes around and is connected with the four upright posts to realize up-and-down sliding, wherein the upright post 2 with holes can realize the positioning function. The silica gel cell culture substrate 13 contains densely distributed magnetic beads, and the magnetic beads can be moved by magnetic force by switching on current to the lower energizing solenoid 3 and the upper energizing solenoid 6, so that the silica gel cell culture substrate 13 is driven to generate upward or downward tensile stress, and an external load is applied.

The working principle of the invention is as follows:

the device is a device for loading of cell engineering cultures and providing a flowing liquid environment. The silica gel cell culture base 13 is embedded with a certain number of densely distributed magnetic beads, when the lower power solenoid 3 and the upper power solenoid 6 on the upper and lower sides of the cell culture chamber 10 are connected with current, a magnetic field is generated, magnetic force is generated on the magnetic beads in the silica gel cell culture base 13, and by adjusting the current between the two power solenoids, the magnetic beads can be attracted to move towards a certain direction, the whole silica gel cell culture base 13 is driven to generate tensile stress, and therefore the tensile stress loading on cells is achieved. In addition, the cell culture chamber 10 has openings on both sides, and can be connected to a pump through a conduit to introduce a cell culture solution, thereby forming a flowing cell culture environment. The invention simulates the mechanical stress on cells under physiological conditions, improves the cell metabolism, promotes the secretion function of the cells, is expected to differentiate the cells in different directions, and is used for other relevant experimental researches of cell engineering.

The invention has the advantages that:

the invention simulates the mechanical stress on cells under physiological conditions, can be loaded by using loads with different sizes, has simple structure, convenient use and wide application, and realizes the continuous and constant loading of the cell culture membrane by adjusting the current in the electrified solenoid. The loading external force can be automatically controlled, and the loading process is simple and easy to operate. The device is small in size and convenient to store. The cost is low, and a plurality of comparison tests can be carried out.

Drawings

FIG. 1 is a schematic view of the structure of the dynamic cell culture apparatus.

FIG. 2 is a schematic diagram showing the structure of a cell culture section of the dynamic cell culture apparatus.

FIG. 3 is a schematic circuit configuration diagram of the dynamic cell culture apparatus.

FIG. 4 is a schematic diagram showing the deformation of the silica gel cell culture substrate of the dynamic cell culture apparatus.

FIG. 5 is a schematic view of the positioning structure of the cell culture apparatus of the dynamic cell culture apparatus.

In the figure: the device comprises a device base 1, a vertical column 2 with holes, a lower electrified solenoid 3, a nut 4, a bolt 5, an upper electrified solenoid 6, a top cover 7, a non-porous vertical column 8, a culture cavity cover 9, a cell culture cavity 10, a culture substrate upper clamping cover 11, a sealing rubber ring 12, a silica gel cell culture substrate 13, a culture substrate lower clamping cover 14 and a positioning pin 15.

Detailed Description

Installation description:

firstly, the four upright posts are fixed on a device base 1 through screws, wherein the upright post with holes 2 and the upright post without holes 8 are arranged in a diagonal line, then the lower electrified solenoid 3 is fixed inside a positioning groove at the center of the device base 1, holes around the cell culture cavity 10 are aligned with the four upright posts and are placed in the four upright posts, and at the moment, the cell culture cavity 10 can slide up and down in the device. And then the upper electrified solenoid 6 is fixed inside a positioning groove at the center of the top cover 7 by the same method, and finally the top cover 7 and the upper electrified solenoid 6 are connected with four upright posts together by screws, thus finishing the installation of the fixed part of the device.

Before use, the cell culture chamber 10 is slid to the highest position and fixed by the positioning pin 15. And then clamping the silica gel cell culture substrate 13 between the upper clamping cover 11 and the lower clamping cover 14 of the culture substrate through four bolts, planting cells on the clamped silica gel cell culture substrate 13, screwing the whole body below the center of the cell culture cavity 10 through threads, and covering the culture cavity cover 9. Thereafter, the position of the cell culture chamber 10 is adjusted and the fixing is performed again. Thus, the installation of the whole force application device is completed.

Cell culture solution is pumped into the cell culture cavity 10 through a pump, when the liquid environment of cell culture is stable, the upper electrified solenoid 6 and the lower electrified solenoid 3 are respectively connected into a circuit, and the deformation conditions of the silica gel cell culture substrate 13 can be observed to be respectively convex, horizontal and concave through adjusting the difference of the relative magnitude of current in the upper electrified solenoid 6 and the lower electrified solenoid 3. Thus, a constant applied load is realized, the cells are in a specific mechanical environment, and the cells can be cultured and observed.

Claims

1. The invention relates to a dynamic culture device for applying a stress/strain gradient field to cells by using magnetic force, which comprises the following components: the device comprises a device base, an electrified solenoid, a nut, a bolt, a top cover, a column with a hole, a column without a hole, a culture cavity cover, a cell culture cavity, a clamping cover above a culture substrate, a sealing rubber ring, a silica gel cell culture substrate, a clamping cover below the culture substrate and a positioning pin.

2. The dynamic cell culture device according to claim 1, wherein the silica gel culture substrate is disposed at the bottom of the cell culture chamber and is fastened by the upper and lower clamping covers through threads, and the clamping cover above the culture substrate has threads that are engaged with the threads of the inner ring below the cell culture chamber and can be removed from the bottom of the cell culture chamber independently.

3. The dynamic cell culture device according to claim 1, wherein the silica gel cell culture substrate is silica gel and contains dense magnetic beads therein, and the magnetic beads are displaced by magnetic force to stretch and deform the silica gel cell culture substrate.

4. The dynamic cell culture apparatus as claimed in claim 1, wherein the electric solenoid is fixed to the base and disposed above and below the cell culture chamber to provide magnetic force from two directions.

5. The dynamic cell culture device according to claim 1, wherein the cell culture chamber is provided with small holes at both ends for connecting culture solution delivery pipes.

6. The dynamic cell culture device according to claim 1, wherein the cell culture chamber is made of polystyrene.

7. The dynamic cell culture device according to claim 1, wherein the upright, the cell culture chamber, the device base and the top cover are connected by screw threads.

8. The dynamic cell culture apparatus according to claim 1, wherein the cell culture chamber has positioning holes on the diagonal line, and the positioning holes are positioned with respect to the vertical columns having holes and the positioning pins, so that the cell culture chamber can be adjusted in the vertical direction.

9. The dynamic cell culture device according to claim 1, wherein the current applied to the energized solenoid is adjustable to adjust the magnetic force provided by the energized solenoid, thereby further controlling the deformation of the silica gel cell culture substrate.

10. The dynamic cell culture apparatus according to claim 1, wherein the cell culture chamber can be adjusted in up-and-down position to adjust the magnetic force applied thereto, and is easy to install.