Substance and method for splitting induced dry-cell to cartilage

A stem cell differentiation and stem cell technology, which is applied in the fields of medicine and biomedical engineering, can solve the problems of disease transmission and insufficient source of autologous chondrocytes, and achieve the effect of sufficient source, easy large-scale promotion and application, and easy acquisition

Active Publication Date: 2007-09-05
SHANGHAI TISSUE ENG LIFE SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the biggest disadvantage of this chondrocyte/stem cell mixed culture induction method is that chondrocytes must directly participate in the chondrogenic differentiation of stem cells or the construction of tissue engineered ca

Method used

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  • Substance and method for splitting induced dry-cell to cartilage
  • Substance and method for splitting induced dry-cell to cartilage
  • Substance and method for splitting induced dry-cell to cartilage

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0078] The preparation method of inducer

[0079] Separating (such as filtering or centrifuging) the culture solution obtained by culturing chondrocytes or cartilage tissues in vitro to remove the chondrocytes and cartilage tissues, so as to obtain a solution free of cells and cell debris.

[0080] A preferred method is to separate the culture fluid obtained by culturing chondrocytes or cartilage tissues in vitro by centrifugation to remove the chondrocytes and cartilage tissues, thereby obtaining a filtrate free of cells and cell debris.

[0081] The culture medium is the culture medium obtained during culturing chondrocytes or cartilage tissues for 1-200 days; more preferably, the time for culturing chondrocytes is 1 day-90 days, and the time for cultivating tissue-engineered cartilage constructed by chondrocytes is 3 days - 180 days.

[0082] The culture fluid is the culture fluid obtained during 24 hours to 150 hours after the liquid change; more preferably, when the cult...

Example Embodiment

[0108] Example 1

[0109] Preparation of inducer I for inducing stem cells to differentiate into cartilage

[0110] 1. Isolation and culture of chondrocytes

[0111] Experimental animals:

[0112] 5 young pigs, weighing about 10kg, male or female.

[0113] experimental method:

[0114] 1. Animals were anesthetized with ketamine 10-20 mg / kg and atropine 0.5-1 mg intramuscularly to induce and inhibit glandular secretion, and slowly push 1 ml / kg of chloral hydrate (10%) or sodium pentobarbital ( 2.5%) 1ml / kg anesthetized;

[0115]2. Routinely disinfect the drape, aseptically cut the knee articular cartilage tissue, cut it into 2mm×3mm pieces, and wash it twice with phosphate buffer;

[0116] 3. According to the modified Klagsbrun method, add 5 times the volume of 1 mg / ml type II collagenase (Worthington, Freehold, NJ, USA), place it in a constant temperature shaker at 37°C for 8-12 hours, filter, and centrifuge;

[0117] 4. The precipitated cells were washed twice with PBS,...

Example Embodiment

[0124] Example 2

[0125] Preparation of Inducer II for Inducing Stem Cell Differentiation to Cartilage

[0126] 1. Chondrocyte-material composite construction

[0127] (1) Preparation of polyglycolic acid (PGA) three-dimensional scaffold

[0128] Non-woven PGA fibers were purchased from Albany Company (Albany, NY, USA) and stored in vacuum. The fiber diameter is 13-15μm, and the PGA fiber is accurately weighed to 20mg / piece, and is pressed into small cylindrical pieces with a diameter of 13mm and a thickness of 2mm with a special mold. After the shape is fixed, the material support is completely immersed in 75% ethanol Disinfect for 30 minutes. Rinse with PBS 3 times, then soak in DMEM medium containing 10% fetal bovine serum for 10 minutes, blot dry and prepare to inoculate cells.

[0129] (2) Construction of chondrocyte-material complex

[0130] 1. When the first-generation chondrocytes cultured in monolayer reached 90% confluence, they were digested with 0.25% trypsin...

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Abstract

A substance for inducing stem-cell to differentiate cartilage and its method are disclosed. It can induce stem-cell into cartilage cell or stem-cell-degradable material composite effectively to form into tissue engineering cartilage. It's safe, economical, convenient and efficient.

Description

technical field [0001] The present invention relates to the fields of medicine and biomedical engineering, more specifically to a method for preparing chondrocytes or cartilage tissues and / or soluble substances secreted by both, and a method and application for inducing stem cells to differentiate into cartilage. Background technique [0002] Defects or damage to cartilage can be caused by congenital malformations, trauma, infection, tumors, or diseases such as osteochondritis. Due to the low self-repair ability of cartilage tissue after injury, the treatment of cartilage defect or injury has always been a great challenge in reconstructive surgery. At present, the commonly used clinical treatment methods include: free or pedicled autologous cartilage transplantation (mainly taken from costal cartilage), free transplantation of allogeneic cartilage, autologous or allogeneic chondrocyte injection transplantation, periosteum or perichondrium transplantation, and artificial pros...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/077
Inventor 周广东曹谊林刘霞刘伟崔磊
Owner SHANGHAI TISSUE ENG LIFE SCI
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