WO2016054847A1 - Structure bionique contenant des canaux et dispositif d'entrainement par force électromagnétique et procédé associé - Google Patents

Structure bionique contenant des canaux et dispositif d'entrainement par force électromagnétique et procédé associé Download PDF

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WO2016054847A1
WO2016054847A1 PCT/CN2014/089771 CN2014089771W WO2016054847A1 WO 2016054847 A1 WO2016054847 A1 WO 2016054847A1 CN 2014089771 W CN2014089771 W CN 2014089771W WO 2016054847 A1 WO2016054847 A1 WO 2016054847A1
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channel
cells
cell
pulsation
bionic structure
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PCT/CN2014/089771
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English (en)
Chinese (zh)
Inventor
王小红
许雨帆
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清华大学
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Priority claimed from CN201410535181.8A external-priority patent/CN104306083B/zh
Application filed by 清华大学 filed Critical 清华大学
Publication of WO2016054847A1 publication Critical patent/WO2016054847A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body

Definitions

  • organ transplantation technology has always faced a series of problems such as immune rejection, donor shortage, organ distribution, and ethics. According to statistics, more than 1.5 million patients need organ transplants each year in China, but the supply-demand ratio is less than 1:100 [Langer R. Tissue Engineering, 2007, 13(1): 1-2]. Tissue Engineering and Organ Manufacturing use a combination of engineering and medicine to create a new solution to this problem from the organizational and organ levels.
  • the biomimetic structure of artificial tissue or organ can be used to implant the body to repair tissue defects, replacing organ function; or as an extracorporeal device, temporarily replacing organ function [Cao Yilin et al., Journal of Clinical Surgery, 2007, 15(1): 40- 41].
  • tissue organs constructed by tissue engineering methods have been clinically applied, but these applications are mostly confined to tissues such as cartilage and epithelium. These tissues are simple in structure, single in composition, and limited in size.
  • the main problem is to construct functional multi-organ substitutes while constructing functional tissues and scaffold structures. These functions mainly include circulatory system, nervous system and immune system.
  • a channel-containing biomimetic structure characterized in that the biomimetic structure comprises a structural body and at least one channel, the channel is distributed in the structural body; the structural body is a mixture of a natural polymer hydrogel and a cell.
  • the cell concentration is 10-10 8 /mL, and the cell is at least one of an embryonic stem cell, an adult stem cell, an adult cell, a cancer cell, and an induced pluripotent stem cell;
  • the channel is a single channel or a branch channel, and the channel is One or more combinations of through-holes at both ends, blind holes at both ends, and one-end through-hole structures at one end; the positional relationship between the channels is intersecting, parallel, collinear or different; the outer wall of the channel and the inner wall of the channel Or cells are distributed in the pores of the channel, and the cells are at least two of the seed cells of the nervous system, the seed cells of the vascular system, and the seed cells of the immune system.
  • the cross section of the channel is circular, elliptical, polygonal or irregular geometrical, and the channel cross-sectional area is 100 ⁇ m 2 -1 cm 2 .
  • the channel-containing biomimetic structure is characterized in that: the nervous system seed cells are at least one of neurons and glial cells; and the blood vessel seed cells are endothelial cells, smooth muscle cells and adipose stem cells. At least one of the immune system seed cells is at least one of lymphocytes and innate immune cells.
  • the natural polymer hydrogel of the present invention is at least one of sodium alginate, collagen, matrigel, dextrose, chitosan, gelatin and fibrinogen, and the mass concentration of the hydrogel is 0.1 to 20%.
  • the natural polymer hydrogel is compounded with at least one of a cell cryopreservative, a cell growth factor, a drug, an anticoagulant, and a magnetic nanoparticle;
  • the cell cryopreservative is dimethyl sulfoxide, glycerin And at least one of dextrose;
  • the cell growth factor is at least one of vascular endothelial growth factor, basic fibroblast growth factor, hepatocyte growth factor, human platelet derived growth factor, and transforming growth factor;
  • the drug is at least one of an antitumor drug and a virus vaccine;
  • the anticoagulant is at least one of heparin and paclitaxel;
  • the magnetic nanoparticles are ferrite particles, metal particles and nitriding
  • the present invention provides an electromagnetic force training device for a channel-containing bionic structure, characterized in that the device comprises an electric field generating system, a magnetic field generating system, a sample stage and a fixed platform; the magnetic field generating system and the sample stage are fixed at a fixed position On the platform; the electric field generating system includes a positive electrode and a negative electrode; the sample stage is provided with a guide rail, and the positive electrode and the negative electrode are respectively mounted on the guide rail through a slider; the sample stage is located in the magnetic field generating system; the magnetic field generating system
  • the sub-assembly includes a bracket and a rotatable ring mounted in the hollow structure of the bracket, and a detachable S pole and N pole are radially symmetrically arranged on the rotatable ring.
  • the present invention also provides a method of training a biomimetic structure using an electric field of the above apparatus, characterized in that the method comprises the following steps:
  • the current is greater than 0, less than or equal to 50 mA; the voltage is greater than 0, less than or equal to 50 V
  • the current direction is alternating between AC, DC or both;
  • the bionic structure After completing the training of the electric field on the bionic structure containing the channel, the bionic structure is cryopreserved or continuously cultured in vitro or directly used for organ transplantation.
  • the bionic structure After completing the training of the magnetic field on the channel-containing bionic structure, the bionic structure is cryopreserved or continuously cultured in vitro or directly used for organ transplantation.
  • the present invention also provides a method of training a biomimetic structure using an electromagnetic composite field of the above apparatus, characterized in that the method comprises the following steps:
  • the pulsation system motor is connected to the pulsation system rail-slider mechanism by a crank;
  • the guide rail-slider The mechanism is connected with the culture liquid supply syringe;
  • the bionic structure is connected to the pulsation culture system through the flow guiding tube;
  • the culture liquid is unidirectionally flowed between the draft tube and the bionic structure containing the channel;
  • the liquid flow rate is greater than 0, less than At 30 cm/s, the action of the pulsation-electric field, the pulsation-magnetic field or the pulsation-electromagnetic composite field is performed; after the training of the bionic structure is completed, the bionic structure is cryopreserved, or the in vitro culture is continued or directly used for organ transplantation.
  • the present invention has the following advantages and outstanding technical effects:
  • the invention is based on the existing three-dimensional forming technology such as 3D printing method and rotating combined mold method, and further compounding the cell and polymer gel mixture to prepare a bionic structure of channel, multi-system and multifunctional complex organizer.
  • the channel-containing biomimetic structure encompasses the vascular system, the nervous system, and the immune system, and structurally and functionally mimics the true multi-system state of the body.
  • the electromagnetic force training device of the invention combines the functions of electric field, magnetic field and pulsation culture, and performs in vitro training and mechanical culture on the bionic structure, so that the cells can be positioned layer by layer and arranged in the pipeline structure under the action of the composite field. Peripheral, close to the cell morphology of the real organ duct, the formed structure has good performance in morphology, immunophysiology.
  • an electromagnetic force training device can use an electric field alone, use a magnetic field alone or simultaneously use an electromagnetic field; in addition, the introduction of a pulsation culture system also increases the diversification of device functions.
  • Figure 1 is a schematic diagram of a bionic tissue structure containing channels.
  • FIG. 2a-2i are schematic views of the state of several channel cells; wherein, FIG. 2a is a schematic diagram of a cell located on the outer wall of the channel, FIG. 2b is a schematic view of a cell located on the inner wall of the channel, and FIG. 2c is a view of a cell located in the channel hole.
  • Schematic diagram, Fig. 2d is a schematic diagram of two cells located on the outer wall of the channel
  • Fig. 2e is a schematic diagram of two cells located on the inner wall of the channel
  • Fig. 2f is a schematic diagram of two cells in the channel hole
  • Fig. 2g is a cell located on the outer wall of the channel.
  • 2h is a schematic diagram of a cell located in the channel wall and another cell located in the channel hole.
  • FIG. 2i is a schematic diagram of a cell located on the outer wall of the channel and another cell located on the inner wall of the channel.
  • Fig. 3 is a schematic view showing the formation of a bionic structure containing a channel by a multi-nozzle 3D printing method.
  • 4a and 4b are respectively a schematic view and an exploded view of a bionic structure of a multi-core rotating combined mold forming channel.
  • 5a, 5b and 5c are respectively a schematic view of a spindle-shaped two-channel biomimetic structure, a schematic diagram of a block-shaped cross-channel bionic structure and a block-like parallel channel bionic structure.
  • Figure 6 is a schematic view of an electromagnetic training device.
  • Figure 7a is a schematic diagram of the explosion of the magnetic field generating system
  • Figure 7b is a schematic diagram of the pulsating culture system.
  • Figure 8a shows the bionic structure containing the channel between the metal plate electrodes
  • Figure 8b shows the bionic structure of the channel between the metal wire or the metal probe
  • Figure 8c is a schematic diagram of the movement of the charged particles in the electric field (E).
  • Fig. 9a shows that the bionic structure containing the channel is located between the magnetic field S and the N pole
  • Fig. 9b is a schematic view of the movement of the magnetic particle in the magnetic field (B) (the broken line indicates the direction of the magnetic field rotation).
  • Fig. 10a shows that the bionic structure containing the channel is located between the positive and negative electric fields and the magnetic field S and N
  • Fig. 10b is a schematic diagram of the movement of the charged particles in the composite electromagnetic field (the broken line indicates the direction of rotation of the magnetic field).
  • a channel-containing biomimetic structure includes a structural body 101 and at least one channel distributed in the structural body 101; the structural body is a mixture of a natural polymer hydrogel and a cell.
  • the cell concentration is 10-10 8 /mL, the cell is at least one of an embryonic stem cell, an adult stem cell, an adult cell, a cancer cell and an induced pluripotent stem cell;
  • the channel is a single channel or a branch channel 105, the channel One or more combinations of the through holes 103 at both ends, the blind holes 106 at both ends, and the one end through holes 104 at one end; the positional relationship between the channels is intersecting, parallel, collinear or different;
  • Cells are distributed in the outer wall, the inner wall of the channel or the pores of the channel, and the cells are at least two of the seed cells of the nervous system, the seed cells of the vascular system, and the seed cells of the immune system.
  • the channel has a circular, elliptical, polygonal or irregular geometry and has a channel cross-sectional area of 100 ⁇ m 2 -1 cm 2 .
  • the nervous system seed cell is at least one of a neuron and a glial cell;
  • the blood vessel seed cell is at least one of an endothelial cell, a smooth muscle cell, and an adipose stem cell;
  • the immune system seed cell is a lymphocyte and At least one of innate immune cells.
  • FIG. 2a-2i are schematic views of the state of several channel cells; wherein, FIG. 2a is a schematic diagram of a cell located on the outer wall of the channel, FIG. 2b is a schematic view of a cell located on the inner wall of the channel, and FIG. 2c is a view of a cell located in the channel hole.
  • Schematic diagram Fig. 2d is a schematic diagram of two cells located on the outer wall of the channel, Fig. 2e is a schematic diagram of two cells located on the inner wall of the channel, and Fig. 2f is two thin Schematic diagram of the cell located in the pore of the channel, FIG. 2g is a schematic diagram of another cell located in the channel pore on the outer wall of the channel, and FIG.
  • FIG. 2h is a schematic diagram of another cell located in the channel hole on the inner wall of the channel
  • FIG. 2i A schematic diagram of a cell located on the outer wall of the channel and another cell located on the inner wall of the channel.
  • FIG. 4a and FIG. 4b it is a preparation method of a biomimetic structure.
  • Fig. 3 is a schematic view showing the formation of a bionic structure containing a channel by a multi-nozzle 3D printing method.
  • 4a and 4b are respectively a schematic view and an exploded view of a bionic structure of a multi-core rotating combined mold forming channel.
  • Figures 5a to 5c are schematic views of several biomimetic structures containing multiple channels.
  • 5a, 5b and 5c are respectively a schematic view of a spindle-shaped two-channel biomimetic structure, a schematic diagram of a block-shaped cross-channel bionic structure and a block-like parallel channel bionic structure.
  • the natural polymer hydrogel is at least one of sodium alginate, collagen, matrigel, dextrose, chitosan, gelatin and fibrinogen, and the mass concentration of the hydrogel is 0.1 to 20%. .
  • the natural polymer hydrogel is compounded with at least one of a cell cryopreservative, a cell growth factor, a drug, an anticoagulant, and a magnetic nanoparticle;
  • the cell cryopreservative is dimethyl sulfoxide, glycerin And at least one of dextrose;
  • the cell growth factor is at least one of vascular endothelial growth factor, basic fibroblast growth factor, hepatocyte growth factor, human platelet derived growth factor, and transforming growth factor;
  • the drug is at least one of an antitumor drug and a virus vaccine;
  • the anticoagulant is at least one of heparin and paclitaxel;
  • the magnetic nanoparticles are ferrite particles, metal particles and nitriding At least one of the iron particles.
  • an electromagnetic training device includes an electric field generating system 601, a magnetic field generating system, a sample stage 603, and a fixed platform 605; the magnetic field generating system and sample stage 603 are mounted on a fixed platform 605;
  • the system 601 includes a positive electrode and a negative electrode;
  • the sample stage 603 is provided with a guide rail, and the positive electrode and the negative electrode are respectively mounted on the guide rail through a slider;
  • the sample stage 603 is located in the magnetic field generating system;
  • the magnetic field generating system includes a bracket 602 and a rotatable ring 608, the rotatable ring 608 is mounted in the hollow structure of the bracket 602, and the detachable S pole 606 and the N pole 607 are radially symmetrically arranged on the rotatable ring 608.
  • the device may further include a pulsation culture system 701; the pulsation culture system is mounted on the fixed platform 605; the pulsation culture system 701 includes a pulsation system motor 702, a pulsation system guide-slider mechanism 703, a culture fluid supply injector 704, a check valve 705, a draft tube 706 and a culture fluid bottle 708; the pulsation system motor 702 is connected to the pulsation system rail-slider mechanism 703 by a crank; the pulsation system rail-slider mechanism 703 and the culture fluid supply syringe 704 Connection, see Figure 7b.
  • the positive electrode and the negative electrode are one of a metal plate, a metal wire or a metal probe.
  • the S pole and the N are one of a permanent magnet or an electromagnet.
  • FIG. 8a the channel-containing biomimetic structure is located between the metal plate electrodes
  • FIG. 8b shows that the channel-containing bionic structure is located between the metal wires or the metal probes
  • FIG. 8c is a schematic diagram of the movement of the charged particles in the electric field (E). .
  • the method for training a channel-containing biomimetic structure in an electric field of the device includes the following steps: a) immersing the prepared channel-containing biomimetic structure in a cell culture liquid containing suspended cells, and placing the electromagnetic force On the sample stage of the training device; b) starting the electric field generating system of the electromagnetic training device, removing the N and S stages of the magnetic field generating system; and making the bionic structure containing the channel in the electric field; the current is greater than 0, less than or equal to 50 mA; It is greater than 0, less than or equal to 50V; the current direction is alternately used by alternating current, direct current or both; c) after completing the training of the electric field on the bionic structure containing the channel, the biomimetic structure is cryopreserved or continuously cultured in vitro or directly used Organ transplantation.
  • Fig. 9a the channel-containing biomimetic structure is located between the magnetic field S and the N pole
  • Fig. 9b is a schematic diagram of the movement of the magnetic particles in the magnetic field (B) (the broken line indicates the direction of the magnetic field rotation).
  • the method for training a channel-containing biomimetic structure in a magnetic field of the device includes the following steps: a) immersing the prepared channel-containing biomimetic structure in a cell culture liquid containing suspended cells, and placing the electromagnetic force On the sample stage of the training device; b) turning off the electric field generating system of the electromagnetic training device, starting the magnetic field generating system, rotating the S pole and the N pole with the rotatable ring, controlling the magnetic induction intensity to be greater than 0, less than or equal to 5T; c) completing After the magnetic field is trained on the bionic structure containing the channel, the bionic structure is cryopreserved or continuously cultured in vitro or directly used for organ transplantation.
  • Fig. 10a shows that the bionic structure containing the channel is located between the positive and negative electric fields and the magnetic field S and N
  • Fig. 10b is a schematic diagram of the movement of the charged particles in the composite electromagnetic field (the broken line indicates the direction of rotation of the magnetic field).
  • the method for training a channel-containing biomimetic structure in the electromagnetic composite field of the device comprises the following steps: a) immersing the prepared channel-containing biomimetic structure in a cell culture fluid containing suspended cells, and placing On the sample stage of the electromagnetic force training device; b) simultaneously starting the electric field generating system and the magnetic field generating system of the electromagnetic training device, so that the bionic structure containing the channel is located in the electric field and the magnetic field; the control current is greater than 0, less than or equal to 50 mA; and the voltage is greater than 0, less than or equal to 50V; current direction is alternating between alternating current, direct current or both; controlling magnetic induction intensity is greater than 0, less than or equal to 5T; c) after completing the electric field and magnetic field simultaneously training the bionic structure containing the channel, the bionic structure Store at low temperature, or continue in vitro culture or directly for organ transplantation.
  • the present invention also applies the training method to the pulsation culture system, the steps including the above steps, and the introduction and use of the pulsation culture system: starting a pulsation culture system 701, the pulsation culture system 701 includes a pulsation system motor 702, a pulsation system guide-slider mechanism 703, a culture fluid supply syringe 704, a check valve 705, a draft tube 706, and a culture fluid bottle 708;
  • the system motor 702 is coupled to the pulsation system rail-slider mechanism 703 by a crank; the rail-slider mechanism 703 is coupled to the culture fluid supply injector 704;
  • the biomimetic structure is coupled to the pulsation culture system via a flow conduit 706;
  • the liquid flows in a one-way flow between the draft tube 706 and the bionic structure containing the channel; the liquid flow rate is greater than 0, less than 30 cm/s,
  • Example 1 A pseudo-heart structure containing two channels of blood vessels and nervous systems was prepared by multi-nozzle 3D printing, and the pseudo-heart structure was trained in an electric field.
  • cardiomyocytes are used as host cells, mixed with the configured sodium alginate solution, and the sodium alginate hydrogel solution has a mass volume concentration of 0.5%; the mixture is then loaded into a nozzle assembly of the multi-nozzle 3D printing device;
  • vascular endothelial cells and neurons as channel cells; mixing vascular endothelial cells with sodium alginate solution into a second nozzle assembly of a 3D printing device; mixing neurons with cell culture fluid (DMEM), loading Into the third nozzle assembly of the 3D printing device;
  • DMEM cell culture fluid
  • d) computer controlled multi-nozzle 3D printing device different nozzle components, control the formation of myocardial cells / sodium alginate in the main structure; synchronous or asynchronous control of vascular endothelial cells / sodium alginate channel formation, get vascular system channels; synchronization or Asynchronously control the neurons/DMEM, locate them in the channel pores, and obtain the nervous system channels; and cross-link the sodium alginate solution with CaCl2 to obtain a sodium alginate hydrogel, which is stacked layer by layer to obtain the multi-channel double The simulated heart structure of the channel;
  • the electric field generating device of the electromagnetic training device will be energized, and the control voltage is DC 50V;
  • Example 2 A multi-nozzle 3D printing method was used to prepare a pseudo-renal structure containing three channels of blood vessels, nerves and immune system, and the simulated kidney structure was trained in an electric field.
  • the embryonic kidney cells are used as host cells, mixed with the configured fibrinogen solution, and the fibrinogen solution has a mass volume concentration of 3%; the mixture is then loaded into a nozzle assembly of the multi-nozzle 3D printing device;
  • vascular endothelial cells, smooth muscle cells, neurons, and T lymphocytes as channel cells; mixing vascular endothelial cells and smooth muscle cells with sodium alginate solution, and loading them into a second nozzle assembly of the 3D printing device; Mixing with the sodium alginate solution, loading into the third nozzle assembly of the 3D printing device; mixing the T lymphocytes with the sodium alginate solution into the fourth nozzle assembly of the 3D printing device;
  • the electric field generating device of the electromagnetic training device will be energized, and the control current is DC 50 mA;
  • Example 3 A multi-nozzle 3D printing method was used to prepare a liver-like structure containing five channels of blood vessels, nerves, bile ducts, and immune system, and the liver-like structure was trained in a magnetic field.
  • vascular endothelial cells and B lymphocytes as channel cells (both of which have been mixed with Fe 3 O 4 magnetic nanoparticles); mixing vascular endothelial cells with collagen solution and loading the second nozzle assembly of the 3D printing device Mixing B lymphocytes with a collagen solution and loading it into a third nozzle assembly of the 3D printing device;
  • d) computer controlled multi-nozzle 3D printing device different nozzle components, control the formation of the main structure of adipose stem cells / gelatin and hepatocytes / gelatin / fibrinogen, and use thrombin to polymerize fibrinogen; synchronous or asynchronous control of vascular endothelium Cell/collagen channel formation, obtaining two vascular channels; synchronous or asynchronous control of B lymphocyte/collagen channel formation, obtaining immune system channels; using glutaraldehyde solution to crosslink collagen into hydrogel, layer by layer, The multi-channel, three-channel, liver-like structure.
  • Example 4 A pancreatic structure containing one-in-one-out dual-vessel and three-channel nervous system was prepared by a multi-core rotation combined mold method, and the bionic pancreatic tissue was trained in a composite electromagnetic field.
  • the islet ⁇ cells as a host cell, mixed with the configured fibrinogen solution, the mass concentration of the fibrinogen solution in the mixture is 5%;
  • DMEM cell culture medium
  • step islet ⁇ cell/fibrinogen mixture into the inner cavity of the top mold through the macropores, and simultaneously rotating the mold base and the top mold at a rotation speed of 30 r/min, and the mold base and the mold branch core are relatively stationary.
  • the perfused mixture forms a semi-spindle shape, and thrombin is used to convert fibrinogen into a hydrogel during rotation;
  • adipose stem cells/DMEM into one-in-one double-tube of a three-channel biomimetic structure precursor to form a vascular system channel; injecting Schwann cells/DMEM into another channel of the channel biomimetic structure precursor to form The nervous system channel; eventually, a three-channel pancreatic structure is obtained.
  • Example 5 A liver structure containing four channels of blood vessels, bile ducts, nerves and immune system was prepared by a multi-core rotating combination mold method, and the liver structure was trained in a composite electromagnetic field.
  • hepatocytes and adipose stem as host cells, mixed with a configured sodium alginate/gelatin solution, fibrinogen solution, wherein the mixture has a volume concentration of sodium alginate and fibrinogen of 5%; in the mixture Adding 3% by volume of DMSO;
  • endothelial cells a) using endothelial cells, Schwann cells, and neutrophils as channel cells; mixing endothelial cells and Schwann cells with cell culture medium (DMEM); mixing neutrophils with sodium alginate solution;
  • DMEM cell culture medium
  • adipose stem cell/heparin/EGF/PBS suspension into the first channel of a four-channel biomimetic structure precursor to form a vascular system channel; perfusion of Schwann cell PBS suspension into a four-channel biomimetic structure precursor In the second channel, the nervous system channel is formed; the neutrophil/PBS suspension is perfused into the third channel of the four-channel biomimetic structure precursor, and CaCl 2 is cross-linked to form the immune system channel; finally, Four-channel semi-spindle liver structure.
  • the two semi-spindle liver precursors are adhered together with a sodium alginate/gelatin/DMSO solution, and a layer of synthetic high molecular polyurethane solution is sprayed to form a spindle-shaped liver precursor;
  • a spindle-shaped liver precursor structure that is driven by a motor to control the rotation of the magnetic field generating system, the magnitude of the magnetic field, and the action time so that the magnetic induction line also passes;
  • Example 6 A four-channel heart structure containing blood vessels, nerves, and immune system was prepared using a multi-core rotating combination mold method, and the heart structure was trained in an electric field.
  • the cardiomyocytes as a host cell, mixed with the configured fibrinogen solution, the mass concentration of the fibrinogen solution in the mixture is 5%; adding a mass volume concentration of 3% dextrose to the mixture;
  • DMEM cell culture medium
  • the myocardial/fibrinogen/dextrose solution mixture is perfused through the macropores into the inner cavity of the top mold, while the mold base and the top mold are relatively rotated, the rotation speed is 30r/min, and the mold base and the mold branch core are Relatively static, the perfused mixture forms a semi-spindle shape, and thrombin is used to convert fibrinogen to hydrogel during rotation;
  • adipose stem cells/EGF/b-FGF/paclitaxel/DMEM perfusion of adipose stem cells/EGF/b-FGF/paclitaxel/DMEM into the first channel of a four-channel biomimetic structure precursor to form a vascular system channel; perfusion of adipose stem cells and smooth muscle cells/DMEM into a four-channel biomimetic structure
  • a vascular system channel is formed; the neuron/DMEM is perfused into the third channel of the four-channel biomimetic structure precursor to form a nervous system channel; before the lymphocyte/DMEM is perfused into the four-channel biomimetic structure
  • the lymphatic passage of the immune system is formed; finally, a four-channel pseudo-cardiac structure is obtained.
  • the electric field generating device of the electromagnetic training device will be energized, and the current is controlled by a direct current alternating current of 30 mA;
  • Example 7 An artificial skin structure containing multiple channels of blood vessels, nerves, and immune system was prepared using a multi-channel detachable combination mold method, and the skin structure was trained in an electromagnetic field.
  • the fibroblasts as a host cell, mixed with the configured sodium alginate solution, the mass concentration of the sodium alginate solution in the mixture is 5%;
  • fibroblasts adipose stem cells as host cells, neurons and lymphocytes as channel cells, keratinocytes, epithelial cells as composite cells, respectively mixed in cell culture medium (DMEM);
  • the electric field generating device of the electromagnetic training device will be energized, and the control current is AC 20 mA;
  • the adipose stem cells as a host cell, mixed with the configured sodium alginate solution, the mass concentration of the sodium alginate solution in the mixture is 1%;
  • endothelial cells a) using endothelial cells, Schwann cells, and neutrophils as channel cells; mixing endothelial cells and Schwann cells with cell culture medium (DMEM); mixing neutrophils with sodium alginate solution;
  • DMEM cell culture medium
  • adipose stem cell/sodium alginate/DMSO solution mixture is poured into the inner cavity of the top mold through the macropores, and the mold base and the top mold are relatively rotated at a rotation speed of 30 r/min, and the mold base is opposite to the mold core. Static, so that the perfused mixture forms a semi-spindle shape, and the sodium alginate is converted into a hydrogel by using CaCl 2 during the rotation;
  • the two semi-spindle breast precursors are adhered together with a fibrinogen/adipose stem cell solution, and a layer of synthetic high molecular polylactic acid and polyglycolic acid copolymer (PLGA) solution is sprayed to form a spindle-shaped breast precursor.
  • PLGA synthetic high molecular polylactic acid and polyglycolic acid copolymer
  • the electric field line passes through the spindle-shaped breast precursor structure at this time;
  • a spindle-shaped breast precursor structure that controls the rotation of the magnetic field generating system, the magnitude of the magnetic field, and the action time by the motor so that the magnetic induction line also passes;
  • Example 9 A lung lobe structure containing two channels of blood vessels, nerves, and immune system was prepared by a combined mold method, and the lung lobe structure was trained in a magnetic field.
  • Adipose stem cells and lung epithelial cells are used as host cells, and mixed with the prepared gelatin solution, the mass concentration of the gelatin solution is 10%, respectively, and the cell cryopreservative glycerin is added in a mass fraction of 3%, and then the mixture is added.
  • Injection To the corresponding position of the combined mold, then inject or infuse the 1% collagen solution onto the corresponding cell layer, freeze the cell gelatin and collagen layer at 37 ° C for 30 minutes, or convert the collagen into a gel using a glutaraldehyde solution;
  • vascular endothelial cells and B lymphocytes as channel cells (both of which have been mixed with Fe 3 O 4 magnetic nanoparticles); mixing vascular endothelial cells with fibrinogen solution and injecting into the two-in and two-out vascular system Mixing B lymphocytes with fibrinogen solution and injecting into a branching system; mixing Schwann cells with fibrinogen solution into another branching nervous system;
  • Fibrinogen was hydrogeled with 0.1% thrombin solution to obtain a four-channel lobes structure.
  • Example 10 A liver-like tumor structure containing a vascular and a nervous system dual channel was prepared by a multi-nozzle 3D printing method, and the liver-like tumor structure was subjected to pulsation culture training in an electric field.
  • liver cancer cells are used as host cells, and mixed with the configured sodium alginate solution, the sodium alginate hydrogel solution has a mass volume concentration of 3%; the mixture is then loaded into a nozzle assembly of the multi-nozzle 3D printing device;
  • vascular endothelial cells and neurons as channel cells; mixing vascular endothelial cells with sodium alginate solution into a second nozzle assembly of a 3D printing device; mixing neurons with cell culture fluid (DMEM), loading Into the third nozzle assembly of the 3D printing device;
  • DMEM cell culture fluid
  • Example 11 A multi-nozzle 3D printing method was used to prepare a pseudo-chondral structure containing three channels of blood vessels, nerves and immune system, and the pseudo-chondral structure was trained in an electric field.
  • the chondrocytes are used as host cells, mixed with the configured fibrinogen solution, and the fibrinogen solution has a mass volume concentration of 3%; the mixture is then loaded into a nozzle assembly of the multi-nozzle 3D printing device;
  • vascular endothelial cells, smooth muscle cells, neurons, and T lymphocytes as channel cells; mixing vascular endothelial cells and smooth muscle cells with sodium alginate solution, and loading them into a second nozzle assembly of the 3D printing device; Mixing with the sodium alginate solution, loading into the third nozzle assembly of the 3D printing device; mixing the T lymphocytes with the sodium alginate solution into the fourth nozzle assembly of the 3D printing device;
  • the electric field generating device of the electromagnetic training device will be energized, and the control current is 2 mA, alternating current;

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  • Health & Medical Sciences (AREA)
  • Cardiology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Transplantation (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Vascular Medicine (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Materials For Medical Uses (AREA)

Abstract

La présente invention concerne une structure bionique contenant des canaux ainsi qu'un dispositif d'entraînement par force électromagnétique et un procédé associé. Une structure bionique en trois dimensions, à fonctions multiples et systèmes multiples contenant des canaux est entraînée in vitro par le dispositif d'entraînement par force électromagnétique. Le corps principal structural du canal est dans un état avec deux trous traversants dans deux extrémités, un état avec deux trous borgnes dans deux extrémités ou un état avec un trou borgne dans une extrémité et un trou traversant dans l'autre extrémité, et des canaux se croisent, parallèles, colinéaires ou bifaciaux les uns par rapport aux autres. La structure bionique dans la présente invention est entraînée ou soumise à un entraînement par pulsations dans des champs électriques, magnétiques ou composites, et les cellules sont orientées en couches disposées dans le canal microfluidique. La structure bionique comprend au moins un type de cellules. Au moins un type de cellules est distribué dans la paroi extérieure, la paroi intérieure ou un trou du canal. Le corps principal (101) de la structure bionique est un mélange de cellules et d'hydrogel polymère naturel. La structure bionique est une substitution pour un tissu ou un organe d'un corps humain, et peut être utilisée pour fournir un support pour le criblage de médicament à rendement élevé, et pour fournir la possibilité d'une transplantation d'organe.
PCT/CN2014/089771 2014-10-11 2014-10-29 Structure bionique contenant des canaux et dispositif d'entrainement par force électromagnétique et procédé associé WO2016054847A1 (fr)

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CN201410535181.8 2014-10-11
CN201410535181.8A CN104306083B (zh) 2014-10-11 一种含通道的仿生结构及其电磁力训练装置和方法

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114274434A (zh) * 2021-12-17 2022-04-05 中国科学院宁波材料技术与工程研究所 一种仿生皮肤的制备装置及其制备方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1237914A (zh) * 1996-11-20 1999-12-08 清水庆彦 人工神经管
GB2386841A (en) * 2002-03-11 2003-10-01 Ind Tech Res Inst Multi-channel bioresorbable nerve regeneration conduit and process for preparing the same
CN1593354A (zh) * 2004-06-25 2005-03-16 清华大学 一种神经组织工程管状支架及其制备方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1237914A (zh) * 1996-11-20 1999-12-08 清水庆彦 人工神经管
GB2386841A (en) * 2002-03-11 2003-10-01 Ind Tech Res Inst Multi-channel bioresorbable nerve regeneration conduit and process for preparing the same
CN1593354A (zh) * 2004-06-25 2005-03-16 清华大学 一种神经组织工程管状支架及其制备方法

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114274434A (zh) * 2021-12-17 2022-04-05 中国科学院宁波材料技术与工程研究所 一种仿生皮肤的制备装置及其制备方法
CN114274434B (zh) * 2021-12-17 2024-02-02 中国科学院宁波材料技术与工程研究所 一种仿生皮肤的制备装置及其制备方法

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