CN-122012232-A - Special low-damage electrotransformation instrument applicable to NK cells and T cells and preparation method thereof

CN122012232ACN 122012232 ACN122012232 ACN 122012232ACN-122012232-A

Abstract

The invention provides a low-damage electric rotating instrument special for NK cells and T cells, wherein an electric rotating box is arranged on an electric rotating instrument body, a fixing groove is formed in an inner cavity of the electric rotating box, an electric rotating cup is arranged on the fixing groove, the electric rotating cup comprises a cup cover, a cup body and an electrode, the cup body is of a cavity structure, the electrode comprises an upper arc electrode and a lower arc electrode, the upper arc electrode is arranged on the cup cover, the lower arc electrode is arranged in the bottom end of the cup body of the electric rotating cup, a knob adjusting device which is elastically connected with the middle of the upper arc electrode is further arranged on the cup cover of the electric rotating cup, and the knob adjusting device adjusts the curvature of the upper arc electrode. The invention solves the problems of reduced electric transformation rate and cell survival rate caused by uneven electric field distribution, can control the apoptosis rate of immune cells to be less than or equal to 10 percent after electric transformation, effectively retains the cell activity, and can be widely applied to the fields of immune cell gene modification and tumor immunotherapy.

Inventors

XU ZHIFENG
LIU GUIBO
ZHANG XIN
MA RUBIAO

Assignees

广州沙艾生物科技有限公司

Dates

Publication Date: 20260512
Application Date: 20260130

Claims (10)

1. The low-damage electrotransformation special electrotransformation instrument suitable for NK cells and T cells is characterized in that an electrotransformation box is arranged on the electrotransformation instrument body, and a fixing groove is formed in an inner cavity of the electrotransformation box; an electric rotating cup is arranged on the fixed groove and comprises a cup cover, a cup body and an electrode, wherein the cup body is of a cavity structure; The electrode comprises an upper arc electrode and a lower arc electrode, the upper arc electrode is arranged on the cup cover, and the lower arc electrode is arranged in the bottom end of the cup body of the electric rotating cup; The cup cover of the electric rotating cup is provided with electrode connecting parts along the two ends of the diameter direction of the cup cover, the electrode connecting parts are internally provided with gap grooves, the two ends of the upper arc-shaped electrode are respectively and slidably connected with the electrode connecting parts of the cup cover, the length of the upper arc-shaped electrode exceeds the distance between the two electrode connecting parts of the cup cover, and the two end parts of the upper arc-shaped electrode are provided with allowance lengths which are accommodated in the gap grooves; The cup cover of the electric rotating cup is also provided with a knob adjusting device which is elastically connected with the middle part of the upper arc electrode, and the knob adjusting device adjusts the curvature of the upper arc electrode.
2. The low-damage electrotransformation special-purpose electrotransformation instrument for NK cells and T cells according to claim 1, wherein two ends of the upper arc-shaped electrode are provided with clamping pieces perpendicular to the ends of the arc-shaped electrode, the slit groove is provided with a slit opening, and the length of the clamping pieces is larger than the width of the slit opening of the slit groove.
3. The low-damage electric rotating special instrument for NK cells and T cells according to claim 1, wherein the knob adjusting device comprises an adjusting knob, a knob scale, the adjusting knob is provided with a screw rod and a screwing part fixed on the screw rod, the screw rod of the adjusting knob is in threaded installation with a cup cover of the electric rotating cup, the bottom of the screw rod of the adjusting knob is exposed and abuts against the middle point of an upper arc electrode, and the upper arc electrode is made of elastic conductive materials.
4. The low-damage electrotransport device for use in NK cells and T cells according to claim 1, wherein the inner cavity of the electrotransport cassette is provided with n x n fixed slots arranged in a matrix, n being a natural number not less than 2.
5. The low-damage electrotransport device special for electrotransport of NK cells and T cells according to claim 1, wherein the width of the cup body of the electrotransport cup is 2-4 mm, the cup body of the electrotransport cup is a cube, the electrode is an aluminum sheet electrode, and the material of the electrotransport cup is polycarbonate or quartz.
6. The low-damage electric rotating instrument special for NK cells and T cells according to claim 1, wherein the electric rotating instrument comprises a shell and a sealing cover, the shell adopts a square box body structure, the upper end of the shell is provided with an opening, the sealing cover is matched with the upper end opening of the shell, and a sealing rubber sleeve is arranged on the bottom surface of the sealing cover.
7. The method of using a low-damage electrotransport device for NK cells and T cells according to any one of claims 1 to 6, comprising the steps of: (1) Preparing a cell fluid to be treated, taking the cell fluid to detect the surface tension of the cell fluid, and calculating the curvature or the curvature radius of the cell fluid according to the poplar-Laplace; (2) Calculating the height of the concave surface of the cell liquid in the electric rotating cup according to the curvature or the curvature radius of the cell liquid in the step (1) and the width of the electric rotating cup; (3) Adding the prepared cell liquid to be treated into an electric rotating cup, covering an electric rotating cup cover, and adjusting a knob on the adjusting cup cover to ensure that the height of a concave surface of an upper arc-shaped electrode is consistent with the height of a concave surface of the cell liquid in the electric rotating cup, wherein the calculated concave surface is used for containing the cell liquid; (4) And starting the electrotransfection instrument to carry out electrotransfection.
8. The method of claim 7, wherein the method for measuring the surface tension of the cell fluid by taking the cell fluid is a capillary rising method, and the surface tension, the fluid density, the capillary inner diameter, the liquid level rising height and the contact angle of the cell fluid are measured.
9. The electric rotating cup is characterized by comprising a cup cover, a cup body and an electrode, wherein the cup body is of a cavity structure; The electrode comprises an upper arc electrode and a lower arc electrode, the upper arc electrode is arranged on the cup cover, and the lower arc electrode is arranged in the bottom end of the cup body of the electric rotating cup; The cup cover of the electric rotating cup is provided with electrode connecting parts along the two ends of the diameter direction of the cup cover, the electrode connecting parts are internally provided with gap grooves, the two ends of the upper arc-shaped electrode are respectively and slidably connected with the electrode connecting parts of the cup cover, the length of the upper arc-shaped electrode exceeds the distance between the two electrode connecting parts of the cup cover, and the two end parts of the upper arc-shaped electrode are provided with allowance lengths which are accommodated in the gap grooves; The cup cover of the electric rotating cup is also provided with a knob adjusting device which is elastically connected with the middle part of the upper arc electrode, and the knob adjusting device adjusts the curvature of the upper arc electrode.
10. The electric rotating cup according to claim 9, wherein two ends of the upper arc electrode are provided with clamping pieces perpendicular to the ends of the arc electrode, the slit groove is provided with a slit opening, and the length of the clamping pieces is larger than the width of the slit opening of the slit groove; The knob adjusting device comprises an adjusting knob, a knob scale and a knob handle, wherein the adjusting knob is provided with a screw rod and a hand twisting part fixed on the screw rod, the screw rod of the adjusting knob is in threaded installation with a cup cover of the electric rotating cup, and the bottom of the screw rod of the adjusting knob is exposed and props against the middle point of the upper arc electrode; The width of the cup body of the electric rotating cup is 2-4 mm, the cup body of the electric rotating cup is a cube, the electrode is an aluminum sheet electrode, and the material of the electric rotating cup is polycarbonate or quartz.

Description

Special low-damage electrotransformation instrument applicable to NK cells and T cells and preparation method thereof Technical Field The invention relates to the field of cell biological experimental equipment, in particular to a special electrotransformation instrument for low-damage electrotransformation of NK cells and T cells and a preparation method thereof. Background The traditional cell electrotransfection instrument has the problem that the electric field distribution is uneven, so that the effect of cell electrotransfection is affected. The electrode cup design of the traditional cell electrotransfer instrument has the defect that the cell suspension is in a concave liquid level (high edge and low center) due to the surface tension, so that the contact distance between the edge of the electrode and the liquid level is uneven. The edge resistance is low, the divided voltage is smaller, the cell is difficult to electrically transfect, the central concave surface resistance is high, the voltage is too high, and the cell is easy to be "electrocuted". When the voltage is set for the operation of the electrotransport device, if the voltage is increased so that the edge cells realize perforation, the cell death rate at the concave surface of the center is increased. If the voltage is lowered to reduce the cell death rate, the marginal cells cannot realize perforation, and the electrotransfection efficiency is reduced. Disclosure of Invention The invention aims to overcome the defects of the prior art and provide a low-damage electrotransport converter special for NK cells and T cells and a preparation method thereof. In order to achieve the aim, the technical scheme adopted by the invention is that the special electric converter for low-damage electric conversion of NK cells and T cells is characterized in that an electric converter body is provided with an electric converter box, and the inner cavity of the electric converter box is provided with a fixed groove; an electric rotating cup is arranged on the fixed groove and comprises a cup cover, a cup body and an electrode, wherein the cup body is of a cavity structure; The electrode comprises an upper arc electrode and a lower arc electrode, the upper arc electrode is arranged on the cup cover, and the lower arc electrode is arranged in the bottom end of the cup body of the electric rotating cup; The cup cover of the electric rotating cup is provided with electrode connecting parts along the two ends of the diameter direction of the cup cover, the electrode connecting parts are internally provided with gap grooves, the two ends of the upper arc-shaped electrode are respectively and slidably connected with the electrode connecting parts of the cup cover, the length of the upper arc-shaped electrode exceeds the distance between the two electrode connecting parts of the cup cover, and the two end parts of the upper arc-shaped electrode are provided with allowance lengths which are accommodated in the gap grooves; The cup cover of the electric rotating cup is also provided with a knob adjusting device which is elastically connected with the middle part of the upper arc electrode, and the knob adjusting device adjusts the curvature of the upper arc electrode. Above-mentioned electrode that is applicable to low damage electricity that NK cells and T cells changes special electric transfer appearance's electrode sets up to arc electrode and lower arc electrode, goes up arc electrode's both ends and is connected at the electrode connecting portion of bowl cover respectively in sliding connection, goes up arc electrode length and surpasss the distance of two electrode connecting portions of bowl cover, goes up arc electrode both ends tip and has surplus length to hold in the gap groove. The length of the upper arc electrode exceeds the distance between two electrode connecting parts of the cup cover, the residual length at the two end parts of the upper arc electrode is contained in the gap groove, so that the curvature of the upper arc electrode can be adjusted, when the curvature is increased, the width of the electric rotating cup is consistent, the length of the upper arc electrode leaked out is prolonged, and the length of the residual length at the two end parts of the upper arc electrode contained in the gap groove is shortened. The curvature of the upper arc-shaped electrode is regulated by the knob regulating device to be consistent with the concave curvature of the cells to be tested, so that the problem that cell suspension is in a concave liquid surface (edge is high and center is low) due to surface tension is better avoided, the electrode distances of the cells to be treated and electrically transfected at the edge and the center tend to be consistent, the electric field distribution is more uniform, and the technical problems that when the electrotometer is operated and set with voltage, if the voltage is regulated to be higher, the edge cells re