WO2021208012A1

WO2021208012A1 - Tissue dissociation tube and instrument

Info

Publication number: WO2021208012A1
Application number: PCT/CN2020/085115
Authority: WO
Inventors: Yu Bao
Original assignee: Suzhou Singleron Biotechnologies, Ltd.
Priority date: 2020-04-16
Filing date: 2020-04-16
Publication date: 2021-10-21

Abstract

A tissue dissociation tube (105) and a tissue dissociation instrument (100) are disclosed. The tissue dissociation tube (105) includes: a pressing block for cutting (210), a top sleeve (209), a blade (205), a blade carrier (206), a spiral stir block (207), a spiral stir block sleeve (204), a grinding block (208), a grinding block sleeve (203), a mesh (202) and a centrifugation tube (201). The tissue dissociation instrument (100) includes: a base (101), a control unit, a power unit, a heating unit (104), an operation unit and a power supply unit (102). The instrument (100) can be used to fully automate tissue dissociation process to generate a single cell suspension that can be used directly for downstream analysis.

Description

TISSUE DISSOCIATION TUBE AND INSTRUMENT

TECHNICAL FIELD

The embodiments of the present disclosure relate to, but are not limited to, the field of analysis of cells and, in particular, to a tissue dissociation tube and instrument.

BACKGROUND

Analysis of cells has been an important element of biological research, clinical testing, and drug development. Recent advances of single cell sequencing and other single cell analysis techniques, as well as cell therapies, all depend on analysis of cells from different tissues.

However, dissociation of tissue samples to single cell suspensions has traditionally been a very tedious manual process, requiring different reagents and intensive optimization for different tissue types, and multiple manual steps such as cutting, grinding, enzymatic digestion, and straining. Several commercially available solutions have shortcomings. For example, gentle MACS ^TM Tissue Dissociators (Miltenyi) only enables cutting, grinding, and incubation all in one mix. After that, the blended mixture needs to be manually transferred out of the tube and further processed (e.g. strained to remove cell debris) . This could cause relatively low efficiency and high loss of cells. Optimization for new tissue types is often required.

SUMMARY

In order to overcome the shortcomings of the existing technology, embodiments of the present disclosure provide a tissue dissociation tube and instrument that are more convenient to operate, and more secure to use.

An embodiment of the present disclosure provides a tissue dissociation tube. The tube includes a plurality of components.

Along the central axis of the entire device, the plurality of components includes: a pressing block for cutting; a top sleeve which is used to hold the pressing block so the pressing block could move up and down in the sleeve; a blade; a blade carrier which enable to carry a round of blades (preferably 20 blades) ; a spiral stir block; a spiral stir block sleeve whose edge works with the spiral block’s edge; a grinding block which has a rotating grinding head with radian will; a grinding block sleeve; a mesh and a centrifugation tube.

Optionally, the top sleeve further includes an external connecting unit to connect the centrifugation tube and the type of the centrifugation tube depends on the size of the whole device; a smooth inner wall that allows the pressing block to slide up and down.

Optionally, the pressing block for cutting further includes grooves which matches the blade not only the width and height but also the length; hole in the middle which fits the central shaft of the device.

Optionally, the blade carrier which is connected on the top sleeve further includes a hollow pipe that allows the central axis of the next part to pass through; grooves which matches the blades around the circle. The number and placement angle and placement method of the blades and grooves depend on the needs of the manufacturer.

Optionally, the spiral stir block and its sleeve which is connected on the blade carrier further includes: an upper cavity with a certain angle or radian of the cavity wall in the sleeve, which can make small pieces of tissue or cell masses slide down smoothly; next is the outer edge of the spiral stir block sleeve and the inner edge of the spiral block, they have spiral teeth arranged in circles working together to cut the pieces of tissue into smaller pieces.

Optionally, the grinding block further includes: a smooth outer wall, which can be matched with the sleeve, and his bottom can squeeze the mesh screen, so that the tissue can be ground into single cell, which can be combined with the reagent to form single cell suspension and fall into the centrifuge tube.

An embodiment of the present disclosure further provides a tissue dissociation instrument, which includes a base, a control unit, a power unit, a constant temperature heating unit, an operation unit and a power supply unit. And this tissue dissociation instrument includes the tissue dissociation tube which has been stated above.

BRIEF DESCRIPTION OF DRAWINGS

Figure 1 is the back and left isometric axonometric map of tissue dissociation instrument.

Figure 2 is the front and right isometric axonometric map of tissue dissociation instrument.

Figure 3 is the structure of the tissue dissociation tube without a centrifugation tube in Figure 1.

Figure 4 is an exploded view of the tissue dissociation tube in Figure 1.

Figure 5 is the exploded view of the spiral stir block and its sleeve in Figure 4.

Figure 6 is the structure diagram and sectional view of the spiral stir block and its sleeve in Figure 4.

Figure 7 is the structure diagram of the heating module in Figure 1.

Figure 8 is a sectional view of the heating module in Figure 1.

DETAILED DESCRIPTION

The following is a detailed introduction of the present disclosure in combination with the accompanying drawings and specific embodiments.

As shown in Fig. 1 and Fig. 2, a tissue separator 100 includes a base 101, a power supply module 102, a processor module 103, a heating module 104, a tissue separator 105, a motor 106, and a display screen 107.

The power supply module 102 supplies power to the whole system, including the processor module 103, the heating module 104, the motor 106, and the display screen 107. The user can use the processor module 103 to connect the mouse and keyboard or directly use the display screen 107 with the touch function to control the tissue separator 100, so as to realize the operation and parameter modification of the system. The processor module 103 includes a CPU, a control circuit, a temperature control PID module, a motor driver and other parts, which can realize the control of the whole system.

As shown in Fig. 3 and Fig. 4, the core part of the tissue separator 100 is a tissue detachable tube 105, which comprises: a pressing block for cutting 210; a top sleeve 209; a blade 205; a blade carrier 206; a spiral stir block 207; a spiral stir block sleeve 204; a grinding block 208; a grinding block sleeve 203; a mesh202; and a centrifugation tube 201.

When large pieces of tissue are put into the top sleeve 209, after pressing, being cut, they become tissue blocks, which fall into the spiral stir block 207 and the spiral stir block sleeve 204 together with the separator solution. Here, the tissue will be further crushed into cell blocks less than 1mm. As they enter the grinding block sleeve 203 and the grinding block 208, the rotating grinding head with radian will. The extruded tissue was ground on the mesh 202, and the type of the mesh screen depends on the size of the cell. Finally the single cell suspension fell into the centrifugation tube 201.

As shown in Fig. 5 and Fig. 6, there are a spiral stir block 207; a spiral stir block sleeve 204, They have special shaped teeth on the inner and outer sides, which can cooperate with each other to cut and stir large tissues.

As shown in Fig. 7 and Fig. 8, The specific implementation method of the heating module 104 is as follows: it is divided into three layers, from the inside to the outside: a tube 301 that makes the internal heat evenly, which can be asbestos, ceramics, etc.; a heating tube 302, which can be resistance or semiconductor heater, etc.; and a heat preservation shell 303. A groove is reserved inside the tube 301 to place the temperature probe, and holes are also reserved at the bottom of the heating tube 302 and the heat preservation shell 303 to facilitate the wire of the temperature probe to pass through.

According to the embodiments of the present disclosure, the user can directly dissociate the tissue into a single cell suspension through a preset program without the need for pipetting and manual operation, and perform subsequent centrifugation operation.

Claims

A tissue dissociation tube, comprising:

a plurality of components, which is configured to enable cutting, grinding of a tissue, and straining of a resulting cell mixture to generate a single cell suspension;

wherein each of the plurality of components forms a channel with an internal mechanism and an internal structure along a main axis of the tissue dissociation tube, so that the tissue and a reagent can pass through the channel and be processed to obtain a required single cell suspension.
The tissue dissociation tube according to claim 1, wherein the plurality of components comprises:

a pressing block for cutting;

a top sleeve;

a blade;

a blade carrier;

a spiral stir block;

a spiral stir block sleeve;

a grinding block;

a grinding block sleeve;

a mesh; and

a centrifugation tube.
The tissue dissociation tube according to claim 2, wherein the plurality of components are assembled and connected with each other along a main shaft of the tissue dissociation tube.
The tissue dissociation tube according to claim 3, wherein

the pressing block is configured to be nested in the top sleeve to move up and down;

the blade is configured to be placed in the blade carrier in a circle regularly;

the blade carrier is configured to be connected on the spiral stir block sleeve;

the spiral stir block is configured to be placed in the spiral stir block sleeve, wherein a clearance is disposed between the spiral stir block and the spiral stir block sleeve;

a protruding part at a top of the grinding block is connected with a groove at a bottom of the spiral stir block;

the grinding block has a circular bottom to squeeze the mesh;

the grinding block is configured to rotate around a shaft in the grinding block sleeve, and their sidewalls are close fitting and have no gaps between them; and

the mesh is configured to be connected on the grinding block sleeve outside.
A tissue dissociation instrument, comprising the tissue dissociation tube according to claim 1, wherein the tissue dissociation instrument further comprises:

a base;

a control unit;

a power unit;

a heating unit;

an operation unit; and

a power supply unit.
The instrument according to claim 5, wherein

the base has a suspended bracket to hold the tissue dissociation tube, and several grooves to hold and fix the control unit, the power unit, a constant temperature heating unit, the operation unit and the power supply unit; and there are gaps between them to let a wire go through.
The instrument according to claim 5, wherein

the control unit is configured to be placed on the base and control a whole instrument system, including a motion system, a display system, and a heating system.
The instrument according to claim 5, wherein

the heating unit comprises 3 layers of tubes which are a heat homogenized tube, a heating tube and a heat preservation shell tube.
The instrument according to claim 8, wherein

the heat homogenized tube is placed on the base and wrapped around the tissue dissociation tube, and there is a groove on a wall of the tissue dissociation tube for a sensor being placed.
The instrument according to claim 8, wherein

the heating tube is wrapped around a heat homogenized tube, and there is a hole in a bottom of a pipe for wire.
The instrument according to claim 8, wherein

the heat preservation shell tube is connected on the base and wrapped around the heat homogenized tube, and there is a hole in a bottom of a pipe for wire.