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Preparation method of tissue engineering cell sheet with osteogenic differentiation and vasculogenesis

An angiogenesis and tissue engineering technology, applied in the field of transplantation, can solve the problems of extracellular matrix damage, decreased cell activity, low cell shelf rate and low cell density, and achieves the effect of maintaining coordination, avoiding adverse reactions and improving signal transmission.

Active Publication Date: 2014-05-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Traditional tissue engineering still has many deficiencies in harvesting and transferring cells, which limits the development of tissue engineering
It is difficult for trypsin to control the degree of cell digestion during the process of digesting cells, resulting in damage to the extracellular matrix, decreased cell activity, and increased chances of cell chromosome mutations
The single-cell suspension transplantation method of BMSCs has low cell loading rate and cell density, and it is difficult to form dense target tissues

Method used

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Embodiment Construction

[0010] The present invention is further described below in conjunction with specific embodiment, but protection scope of the present invention is not limited thereto:

[0011] The preparation method of the tissue engineering cell sheet with osteogenic differentiation and angiogenesis of the present invention comprises the following steps:

[0012] 1. Isolation of bone marrow mesenchymal stem cells (MSCs)

[0013] Healthy adult Sprague-Dawley rats were sacrificed after anesthesia, soaked in 75% alcohol for 10 min, and the long bones of the limbs were removed under aseptic conditions, the soft tissue attached to the bone surface was removed, and washed with PBS buffer. The metaphysis at both ends was resected with a rongeur to expose the bone marrow cavity. Thoroughly flush the bone marrow cavity with low-sugar DMEM medium with a volume concentration of 10% FBS, and then repeatedly flush the flushed bone marrow with a syringe with a No. 7 needle and a No. 5 needle to fully ...

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Abstract

The invention discloses a preparation method of a tissue engineering cell sheet with osteogenic differentiation and vasculogenesis. The preparation method comprises the steps of firstly, separating BMSCs (bone marrow mesenchymal stem cells), carrying out passage multiplication culture of the BMSCs, and culturing mesenchymal stem cell sheet to obtain the tissue engineering cell sheet with osteogenic differentiation and vasculogenesis. The cell sheet technology is adopted to separate and culture BMSCs, the obtained BMSCs are of a complete sheet structure containing extracellular matrix and having osteogenic differentiation and vasculogenesis potency. As a tissue formed by pure cells is transplanted into a body, the use of a scaffold and other carriers and the untoward effects are avoided; and besides, the BMSCs cell sheet retains important cell surface protein such as an ion channel, ligandin and the like, intercellular signal transmission can be performed well, the functions are kept to be coordinated, and the cell utilization rate and the cell transferring activity are greatly improved.

Description

technical field [0001] The invention belongs to the field of transplantation, and in particular relates to a preparation method of a cell sheet for tissue repair and regeneration. Background technique [0002] At present, the main method used in tissue engineering is to use degradable biomaterials to make a three-dimensional scaffold similar to the shape of the desired tissue, and to cultivate specific cells on it. After the cells grow into the desired tissue on the scaffold, the tissue and scaffold are combined. Transplanted together into the mother to replace diseased or damaged tissue. Today, scientists have obtained many kinds of tissues through such methods and made great contributions to regenerative medicine, such as the repair of cartilage, bone, and blood vessels, etc. However, the development of tissue engineering also encountered some problems. First, the biocompatibility of the scaffold needs to be verified. Some biological scaffolds can cause inflammatory hype...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61L27/38
Inventor 严伟祺齐义营
Owner ZHEJIANG UNIV
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