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Multi-stage suspension printing method for constructing complex heterogeneous tissue/organ

A tissue and organ technology, applied in the field of multi-level suspension printing, can solve the problems of limited application in the field of translational medicine, limited application, difficult construction, etc.

Active Publication Date: 2021-08-24
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in 2019, based on the suspension printing strategy, the Tal Dvir research group in Israel printed a heart model with vascular structures using bio-inks prepared from extracellular matrix (Noor, N., et al., 3D Printing of personalizedthick and perfusable cardiac patches and hearts .Advanced Science, 2019.6(11):p.190034.); Although this research has achieved the suspension printing of various inks, it is difficult to construct structural features (such as microvessels and nerves) with an accuracy of less than 100 microns, which limits its use in Applications in bionic construction of complex tissues and organs
[0006] In summary, bio-3D printing technology has great advantages in the construction of tissues / organs. However, in vitro construction of tissues / organs with complex vascular channels and heterogeneous cell structures is still a key challenge in the field of regenerative medicine, which greatly limits its application in the field of regenerative medicine. Applications in the field of translational medicine

Method used

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  • Multi-stage suspension printing method for constructing complex heterogeneous tissue/organ
  • Multi-stage suspension printing method for constructing complex heterogeneous tissue/organ
  • Multi-stage suspension printing method for constructing complex heterogeneous tissue/organ

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1, In Vitro Construction of Biomimetic Vascularized Myocardial Chambers

[0053] 1. Preparation of bio-ink (loaded with cardiomyocytes)

[0054] Human-derived induced pluripotent stem cells were cultured in vitro and differentiated to obtain cardiomyocytes and vascular endothelial cells derived from pluripotent stem cells. Photocrosslinkable gelatin methacrylate (GelMA) was used as the microsphere carrier material.

[0055] Using a T-shaped microfluidic device, the density of cardiomyocytes was 1×10 7 The 7.5wt% GelMA solution per mL is passed into the dispersed phase inlet of the T-type microfluidic device, and the flow rate is 0.5mL / h, and the mineral oil containing 10% Span 80 (span 80, surfactant) is passed into the T-type The continuous phase inlet of the microfluidic device has a flow rate of 6.0mL / h, and light crosslinking is carried out at the outlet of the chip to obtain gel microspheres with a diameter of 400 μm to 450 μm ( figure 2 a). By living d...

Embodiment 2

[0062] Embodiment 2, in vitro construction of bionic glioma model

[0063] 1. Preparation of bio-ink (loaded with nerve cells)

[0064] Glioma cells derived from patients are used for in vitro culture and expansion, and human-derived induced pluripotent stem cells are induced to differentiate into nerve cells and endothelial cells, using a coaxial focusing microfluidic device. The nerve cell suspension, the hyaluronic acid solution with a mass fraction of 10.0%, and the Matrigel solution with a volume fraction of 40% (purchased from BD Company) were uniformly mixed in a ratio of 1:2:1, and the final nerve cell density was 2× 10 6 / mL. Pass the hyaluronic acid / Matrigel solution loaded with nerve cells into the dispersed phase inlet of the coaxial focusing microfluidic device at a flow rate of 1mL / h, and pass mineral oil containing 2% Span 80 into the coaxial focusing microfluidic device The continuous phase inlet of the control device, the flow rate is 10.0mL / h, and the gel ...

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Abstract

The invention discloses a multi-stage suspension printing method for constructing a complex heterogeneous tissue / organ. The method comprises the following steps of S1, preparing bio-ink, wherein the bio-ink is formed by cross-linked cell-carrying gel microspheres, or is obtained by mixing the cross-linked cell-carrying gel microspheres with one or more non-cross-linked gel materials; S2, printing the bio-ink in a suspension medium to construct a specific tissue / organ structure; S3, further performing two-stage or multi-stage substructure printing in the tissue / organ structure obtained in S2; and S4, after printing, dissolving out the suspension medium after overall crosslinking. According to the multi-stage suspension three-dimensional printing method, the gel microsphere ink based on shear thinning and self-healing characteristics can be printed and formed in the suspension medium and then can be used as the suspension medium for printing a next-stage structure, is suitable for constructing the tissue and organ model with a blood vessel channel and a heterogeneous cell structure, and promotes the clinical application of the engineered tissue / organ in the aspect of regeneration and repair treatment.

Description

technical field [0001] The invention relates to a multi-level suspension printing method for constructing complex heterogeneous tissues / organs, and belongs to the technical fields of tissue engineering and biomanufacturing. Background technique [0002] Tissue engineering and regenerative medicine, as an emerging interdisciplinary subject, aims to construct artificial tissues and organs in vitro with biomimetic structures and functions. It has broad application prospects in tissue and organ regeneration and repair, drug development and screening, and pathological model construction. At present, tissue engineering products represented by bladder, skin, cartilage and blood vessels have been initially applied. However, the in vitro construction of complex heterogeneous tissues / organs such as the heart and liver is still progressing slowly. [0003] The traditional tissue engineering method mainly adopts a top-down strategy, represented by cell-scaffold composite technology, tha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B29C64/10B29C64/314B29C64/379B29C64/40B33Y10/00B33Y40/10B33Y40/20C12N5/00B29L31/40
CPCB29C64/10B29C64/314B29C64/379B29C64/40B33Y10/00B33Y40/10B33Y40/20C12N5/00B29L2031/40C12N2531/00C12N2535/00
Inventor 熊卓方永聪张婷郭依涵郭昱江
Owner TSINGHUA UNIV
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