JP3163533B2 - Stretch stimulation load device for cultured cells using silicon belt - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a technique for applying a stretching stimulus or a stretching stimulus to cultured cells or tissues.

[0002]

2. Description of the Related Art In general, in order to apply a stretching stimulus to cultured cells and the like, a method has been adopted in which cells are cultured on a silicon film coated with an extracellular matrix or the like, and the cells are stretched together with the silicon film. Conventionally, as a device for extending a silicon film, a unidirectional device (FIG. 4) and a radial device (FIG. 5) have been used in consideration of the direction of extension.

In the case of a unidirectional one, both ends of a rectangular silicon film (9) are tightly fixed using clips (11), and the rotational motion of a synchronous motor using a cam (12) is converted into piston motion. A method of stretching (10) or returning to the original length (9) by a mechanism [conventional method 1] is used [FIG. 4].

[0004] In the case of a radial one, a petri dish (15) having only a bottom surface (13) made of a silicon film is drawn from below to stretch the bottom silicon film (14) or to return to the original state (13). [Conventional method 2] is used (Flexcell Corp., US: Patent No. 47896)
01, 4822741, 4839280) [FIG. 5].

[0005]

[Problems to be solved by the invention]

[Conventional method 1] Since both ends of the silicon film (9) are tightly fixed so that they do not separate even when stretched, a pincushion-like distortion like (10) occurs when the silicon film (9) is stretched. Because the degree of deformation differs depending on the location, it has been difficult to analyze the extension direction. Equipment that includes fixing devices such as clips required sterilization operations.However, there are few springs and screws made of materials that can withstand repeated high-temperature, high-pressure, and high-humidity autoclave sterilization, and are easy to process. Each time the fixing operation was complicated. Also, for devices using synchronous motors and cams, changing the amplitude requires changing the hardware itself, which is troublesome, and it is not possible to set [expansion and retraction including cyclic pause] and change the expansion / contraction acceleration. Met.

[Conventional method 2] Since the stretched silicon film (14) is radially tensioned,
The tension direction differs depending on the location, making it difficult to analyze the extension direction. It was also pointed out that only the peripheral portion of the silicon film was extended, and the central portion was hardly stretched. Furthermore, the extension rate is 2 structurally in the peripheral area.
There was also a restriction that only 4% could be obtained. Since the silicone is stretched by an indirect method of suction, the actual stretching rate may be deviated compared to the set stretching rate due to abrasion of packing or adhesion of dust. In addition, ready-made silicon petri dishes have a fixed size, and thus have a low degree of freedom and are expensive. Further, the suction unit (including the computer and the suction pump) was also large-scale, took up space, and was expensive.

[0007]

In order to solve the above-mentioned problems, the present invention uses a belt-shaped silicon film of a carrier (1) and expands and contracts it using a high-torque stepping motor and a lead screw. [FIG. 1]. Two rods (2) are inserted inside the silicon belt (1) to which cells and the like are attached, and the rods are hooked on the left and right arms (3), respectively. The clockwise and counterclockwise rotational motion of the high torque stepping motor is converted into arm piston motion using a lead screw (4). The silicon belt can be expanded and contracted by swinging the arm left and right.

[0008]

EXAMPLE A silicon belt is formed by a dipping method using a glass test tube or a silicon rod as a template using a sill pot 184W / C manufactured by Dow Corning Asia Co., Ltd., and cured by applying heat. If the stripped bag-shaped silicon is sliced, a silicon belt can be formed.

The silicon belt is autoclaved, and the rod and the arm are dry-heat sterilized independently, but it is also easy to assemble them aseptically.

The rotary motion of the high torque stepper motor was converted to linear motion using a Kirk lead screw.

The rotational motion of the high torque stepping motor can be converted to linear motion using a timing belt.

The movement of the stepping motor of Japan Servo Co., Ltd. can be controlled from 0% to at least 11
The extension rate can be set with a high accuracy of 0.1% increments in the range of 0%, and changes in programs including conversion of expansion / contraction acceleration / extension time / pause / expansion / reduction frequency can be performed without changing sterilized hardware. It was easy to set.

Since the portion of the arm where the rod is hooked needs to be sterilized, it is more convenient to make the movable portion connected to the lead screw detachable using a screw or the like.

When microscopic observation is unnecessary, it is expected that a higher extension rate can be obtained by using latex rubber instead of latex silicon.

[0015]

As described above, the following problems, which have been problems in the conventional apparatus, have been improved.

Since the silicone belt has no seams, it is difficult to cut and the extension rate can be increased (110%,
That is, the extension more than double the original length has been confirmed).

Since the silicon belt is not pressed, there is no distortion of the pincushion type even when the silicon belt is extended (6), and the extension direction and the extension ratio are the same at any position on the belt, making analysis easy [FIG. 2].

By changing the template, the silicon belt can be made in any size at a low price, and the sterilization and the attachment to the device are simplified.

Microscopic observation in a state where the cells are stretched was not possible by the conventional method. However, by inserting glass plates (7) of various lengths into a silicon belt, the cells were stretched at an arbitrary stretching rate. The cells in the state (8) were also observable [FIG. 3].

As the name suggests, a high-torque stepping motor has a strong power, and is a highly accurate motor usually used for positioning a print head of a printer, so that the extension ratio can be set in small steps and over a wide range. . Furthermore, since the silicon belt was stretched directly, the accuracy was high, and there was no room for inconvenience.

The movement of the stepping motor can be easily programmed using a controller, and the extension rate, expansion / contraction acceleration, extension time, pause, expansion / contraction frequency, etc. can be easily changed. By using the present invention, it has become possible for the first time to extend cells over a whole day or to give a stretching stimulus after stretching for a certain period of time.

The movement of the stepping motor can be operated only by the controller program without touching the hardware, and even if the program is changed during expansion and contraction, it can be performed without touching the sterilized parts.

By culturing the cells with the silicon belt extended and returning the cells to the original state, the influence of the shrinkage of the basal plane on the cells can be observed.

It has become possible to conduct experiments by directly connecting a fragment (ring) of a blood vessel to a device.

The extension mechanism using a silicon belt is very simple, and can be simplified in terms of equipment (downsizing) as compared with [Conventional method 2], and the cost of the system is less than 1/10. became.

[Brief description of the drawings]

FIG. 1 is a perspective view of the device of the present invention.

FIG. 2A is a plan view of a silicon belt before expansion and contraction. B is a plan view of the silicon belt after expansion and contraction.

FIG. 3 is a perspective view in which a silicon belt is fixed at the time of extension.

FIG. 4 is a plan view of a unidirectional telescopic device. C is a plan view before expansion and contraction. D is a plan view after expansion and contraction.

FIG. 5 is a perspective view of a radial stretching device. E is a perspective view before expansion and contraction. F It is a perspective view after expansion and contraction.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Silicon belt 2 Rod 3 Arm 4 Lead screw 5 Silicon belt before extension 6 Silicon belt after extension 7 Glass plate 8 Silicon belt fixed in an extended state 9 Silicon film before extension 10 Silicon film after extension 11 Clip 12 Cam 13 Silicon film bottom before extension 14 Silicon film bottom after extension 15 Petri dish (side)

Claims (3)

(57) [Claims] 1. A cultured cell in which a belt-like silicon film (1) is used as a carrier, two rods (2) are inserted into the carrier, and each rod is pulled apart and returned to expand and contract the belt-like silicon film. Telescopic stimulus load device. 2. The method according to claim 1, wherein a stepping motor is used as power, the rotation of the stepping motor is changed to a linear reciprocating movement by a lead screw, and a rod attached thereto is inserted into the silicon belt. 2. The stretch stimulation loading device for cultured cells according to 1. 3. A stepping motor is used as power, and its rotational motion is changed to linear reciprocating motion by a timing belt.
The stretching stimulus load device for cultured cells according to claim 1, wherein the device is realized by inserting a rod (2) attached to the silicon belt (1).