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CN-121975615-A - Single-cell dynamic mechanical loading system and force regulation method based on magnetic beads and bidirectional static magnetic field

CN121975615ACN 121975615 ACN121975615 ACN 121975615ACN-121975615-A

Abstract

The invention belongs to the technical field of cell mechanics, and particularly relates to a single-cell dynamic mechanical loading system and a force regulation method based on magnetic beads and a bidirectional static magnetic field. The invention provides a single-cell dynamic mechanical loading system and a force regulation method based on magnetic beads and a bidirectional static magnetic field. The system realizes single-cell-level pulling/pressing bidirectional mechanical stimulation for the first time, realizes cooperative regulation of magnetic bead concentration and magnetic field direction, breaks through the limitation of the traditional multicellular loading mechanical mode, and can be used for single-cell mechanical response research (such as stem cell differentiation and tumor cell migration), drug screening (for detecting the influence of drugs on cell mechanical characteristics) and construction of dynamic mechanical microenvironment in tissue engineering by mechanically regulating single cells, thereby having good application prospect.

Inventors

  • YU LEIXIAO
  • XIAO LI

Assignees

  • 四川大学

Dates

Publication Date
20260505
Application Date
20251208

Claims (10)

  1. 1. A single-cell dynamic mechanical loading system is characterized by comprising The magnetic field system module is configured to adjust the permanent magnet array according to a mechanical regulation mode, and the magnetic field intensity of the magnetic field of the permanent magnet array at a single cell is 0.5-2T; And the dynamic regulation and control module is configured to regulate the force applied to the single cells by controlling the concentration gradient of the magnetic beads and regulate the direction of the stress of the single cells by the direction of the magnetic field. And the cell stress feedback module is configured to reflect the actual stress of the single cell in real time.
  2. 2. The dynamic mechanical loading system of single cell according to claim 1, wherein the magnetic field intensity of the permanent magnet array magnetic field at the single cell is 1T.
  3. 3. The single-cell dynamic mechanical loading system according to claim 1, wherein the permanent magnet array is arranged above the magnetic field system module when the mechanical regulation mode is a tension mode, and the permanent magnet array is arranged below the magnetic field system module when the mechanical regulation mode is a pressure mode.
  4. 4. The single-cell dynamic mechanical loading system according to claim 1, wherein the concentration of the magnetic beads in the dynamic regulation module is 0-200 mug/mL.
  5. 5. The dynamic mechanical loading system of single cell according to claim 1, wherein the magnetic beads are RGD and polyglycerol co-modified magnetic beads.
  6. 6. The single-cell dynamic mechanical loading system according to claim 5, wherein the RGD and polyglycerol co-modified magnetic beads are prepared by the steps of; Step 1, reacting glycidol with ethyl vinyl ether to prepare ethyl glycidyl ether; Step 2, generating PG-b-PAGE block copolymer by anionic ring-opening copolymerization reaction of ethyl glycidyl ether and allyl glycidyl ether; step 3, coupling the PG-b-PAGE block copolymer with cysteamine hydrochloride through photoinitiated thiol-ene click reaction; step 4, performing condensation reaction on the coupled product and 3, 4-dihydroxyhydrocinnamic acid to obtain catechol functionalized polyglycerol PG-Cat; Step 5, performing cycloaddition reaction on the PG-Cat and the activated RGD to prepare RGD-PG-Cat; and 6, reacting RGD-PG-Cat with amino modified magnetic beads to obtain the magnetic resonance imaging dye.
  7. 7. The dynamic mechanical loading system of single cell according to claim 1, wherein in the cell stress feedback module, the measuring single cell actual stress comprises measuring single cell actual stress according to coulomb's law of magnetic force; in the magnetic field system module, the permanent magnet is selected from neodymium iron boron permanent magnets, the diameter of the magnetic beads in the dynamic regulation module is 1-2 mu m, and the saturation magnetization intensity of the magnetic beads is 3-6 emu/g.
  8. 8. A method for force control using a single cell dynamic mechanical loading system according to any one of claims 1-7, characterized in that it comprises: Step 1, incubating cells and magnetic beads together, and controlling the concentration gradient of the magnetic beads through a dynamic regulation and control module; step 2, setting a mechanical regulation mode; step3, the magnetic field system module adjusts the permanent magnet array; and 4, observing single-cell stress conditions through a cell stress feedback module.
  9. 9. The method of force modulation according to claim 8, wherein in step 1, the concentration of the beads is controlled by a gradient from low to high through a dynamic modulation module; And/or, in the step 4, according to the single-cell stress condition, regulating and controlling the magnetic bead concentration of the dynamic regulating and controlling module.
  10. 10. An apparatus integrated with the single cell dynamic mechanical loading system of any one of claims 1-7.

Description

Single-cell dynamic mechanical loading system and force regulation method based on magnetic beads and bidirectional static magnetic field Technical Field The invention belongs to the technical field of cell mechanics, and particularly relates to a single-cell dynamic mechanical loading system and a force regulation method based on magnetic beads and a bidirectional static magnetic field. Background Cells are in complex mechanical microenvironments in vivo, such as vascular shear forces, tissue compression forces, and the like. These mechanical signals have a direct and profound effect on cell differentiation, migration and disease progression. Therefore, the intensive research of the mechanical response of single cells has extremely important scientific significance and clinical value for understanding the cell behavior mechanism, revealing the disease occurrence and development rule and developing new treatment strategies. However, the existing single-cell mechanical loading technology still has a plurality of limitations, and the requirement of accurately researching single-cell mechanical response is difficult to meet. The conventional single-cell mechanical loading technology mainly comprises the following steps that firstly, a Flexcell cell stretcher applies stretching force to cells through a mechanical device, but the device is usually operated aiming at cell groups, the single cells are difficult to accurately control, and dynamic mechanical loading of the cells cannot be realized. The method for directly pressing the cells by the glass plate can apply pressure to the cells, but the method is rough in operation and low in force application precision, can not realize the bidirectional force loading of pulling/pressing, and is difficult to avoid the mutual interference among multiple cells. In addition, the prior art has the following problems that the force application precision is insufficient, the existing magnetic bead-magnetic field system is large in magnetic field gradient range and cannot focus on single cells aiming at cell groups, the targeting is poor, the magnetic bead and the cells are combined to depend on non-specific adsorption to cause multicellular interference, the mechanical mode is single, a dynamic magnetic field (such as an alternating magnetic field) is easy to generate a thermal effect, the pulling/pressing bidirectional force loading of the magnetic field in one direction is difficult to realize, and the complex mechanical environment is difficult to simulate. These drawbacks severely limit the accurate study of single cell mechanical responses, impeding the development of the field of cell mechanics. Aiming at the defects of the prior art, the development of a precise, dynamic and safe mechanical loading system aiming at single cells is particularly necessary. Disclosure of Invention Aiming at the defects of the prior art, the invention provides a single-cell dynamic mechanical loading system and a force regulation method based on magnetic beads and a bidirectional static magnetic field. The invention provides a single cell dynamic mechanical loading system, which comprises The magnetic field system module is configured to adjust the permanent magnet array according to a mechanical regulation mode, and the magnetic field intensity of the magnetic field of the permanent magnet array at a single cell is 0.5-2T; And the dynamic regulation and control module is configured to regulate the force applied to the single cells by controlling the concentration gradient of the magnetic beads and regulate the direction of the stress of the single cells by the direction of the magnetic field. And the cell stress feedback module is configured to reflect the actual stress of the single cell in real time. Preferably, in the magnetic field system module, the magnetic field strength of the permanent magnet array magnetic field at the single cell is 1T. Preferably, in the magnetic field system module, when the mechanical regulation mode is a tension mode, the permanent magnet array is arranged above the magnetic field system module, and when the mechanical regulation mode is a pressure mode, the permanent magnet array is arranged below the magnetic field system module. Preferably, in the dynamic regulation module, the concentration of the magnetic beads is 0-200 mug/mL. Preferably, the magnetic beads are magnetic beads co-modified by RGD and polyglycerol. Preferably, the RGD and polyglycerol co-modified magnetic beads are prepared according to the following steps; Step 1, reacting glycidol with ethyl vinyl ether to prepare ethyl glycidyl ether; Step 2, generating PG-b-PAGE block copolymer by anionic ring-opening copolymerization reaction of ethyl glycidyl ether and allyl glycidyl ether; step 3, coupling the PG-b-PAGE block copolymer with cysteamine hydrochloride through photoinitiated thiol-ene click reaction; step 4, performing condensation reaction on the coupled product and 3, 4-dihydroxyhyd