Constructing method of bracket-free engineering cartilaginous tissue and product thereof

A construction method and cartilage tissue technology, applied in bone implants, medical science, prostheses, etc., can solve problems such as cumbersome compounding, uneven cell seeding density, compatibility with human body safety, and complicated clinical application approval of scaffolds , to achieve the effect of uniform planting density and simple operation

Inactive Publication Date: 2009-09-30
GENERAL HOSPITAL OF PLA
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  • Abstract
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Problems solved by technology

The compounding of cells and the above-mentioned scaffold materials is cumbersome, the seeding density of cells is uneven, and there are problems such as the bioc

Method used

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  • Constructing method of bracket-free engineering cartilaginous tissue and product thereof
  • Constructing method of bracket-free engineering cartilaginous tissue and product thereof
  • Constructing method of bracket-free engineering cartilaginous tissue and product thereof

Examples

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Embodiment 1

[0034] Example 1 Construction and identification of scaffold-free tissue engineered cartilage tissue

[0035] 1. Isolation and culture of human umbilical cord Wharton's gel stem cells

[0036] Take the umbilical cord tissue of full-term normal and healthy puerpera from the operating table, fully wash the residual blood with D-Hank balance solution, remove the umbilical vein, artery and umbilical cord adventitia, peel off the Wharton glue, shred it, and stir it with type I collagenase at 37°C Digestion under the action of organs. The digested cell-containing digestion solution was washed with D-Hank balance solution, centrifuged at 1500 rpm for 10 min, and centrifuged three times, and the collected cells were planted in DMEM culture flasks with a volume fraction of 10% FBS at 37°C. 5% CO 2 cultured in an incubator and observed with an inverted phase-contrast microscope.

[0037] 2. Detection and identification of molecular markers on the surface of human umbilical cord Whart...

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Abstract

The invention discloses a constructing method of a bracket-free engineering cartilaginous tissue and a product therefrom. The constructing method comprises the following steps: inducing and differentiating human umbilical mesenchymal stem cells into cartilaginous cells and then carrying out aggregation inducing culture so that the cartilaginous cells excrete stroma towards the outside of cells and form membranoid substance; continuously inducing and culturing the membranoid substance in a cell culture box and then transferring the membranoid substance into a centrifugal tube for inducing culture to form loose cartilaginous tissue agglomerate; and continuously inducing and culturing the loose cartilaginous tissue agglomerate in a rotating wall type bioreactor to obtain dense cartilaginous-like tissue. The cytology, the histology or the immunohistochemistry dyeing shows that the constructed engineering tissue has a normal cartilaginous cell type. The constructed bracket-free engineering cartilaginous tissue does not relate to the composite problem of the cells and an exogenous bracket, is absent from the biocompatibility problem between the cells and the bracket, is safe to human body, has even cell planting density and simple clinical application operation and can be applied to the treatment or the repair of cartilage loss.

Description

technical field [0001] The invention relates to a method for constructing engineered tissue, in particular to a method for constructing unsupported engineered cartilage tissue and its products, and belongs to the fields of regenerative medicine and tissue engineering. Background technique [0002] Cartilage injury repair has always been a hot spot in joint surgery research. Autologous chondrocyte implantation (ACI) abroad has achieved good clinical efficacy, but requires a second operation, which increases the suffering of patients. At present, the seed cells used to construct engineered cartilage tissue can be derived from bone marrow mesenchymal stem cells (BMSCs), but the number and proliferation ability of stem cells decrease significantly with age. The use of autologous adipose-derived stem cells in the construction of articular cartilage tissue engineering has also been reported, but these all have the problem of requiring patients to undergo secondary surgery. Embry...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61F2/28
Inventor 卢世璧侯克东袁玫张莉彭江郭金义许文静田玥
Owner GENERAL HOSPITAL OF PLA
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