Method for inducing bone marrow substrate stem cell into cartilage

A technology of stem cells and chondrocytes, applied in the fields of medicine and biomedical engineering, can solve problems such as inability to form cartilage tissue, high cost, and complicated induction methods

Inactive Publication Date: 2007-11-21
上海第二医科大学附属第九人民医院
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Scholars from various countries have conducted a lot of research on MSCs from different tissue sources, and found that MSCs can differentiate into chondrocytes in vitro and express the phenotype and function of chondrocytes, but these induction methods are technically complicated
Moreover, due to the application of a variety of growth factors, the cost is extremely expensive, and the induced MSCs can only repair cartilage defects in the joints, but cannot form or only form a small amount of cartilage tissue in the subcutaneous tissue, and cannot form large pieces. more mature cartilage

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for inducing bone marrow substrate stem cell into cartilage
  • Method for inducing bone marrow substrate stem cell into cartilage
  • Method for inducing bone marrow substrate stem cell into cartilage

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0059] The preparation method of the tissue engineered cartilage of the present invention is simple and convenient, and only needs to mix a certain amount of the mixture of chondrocytes and bone marrow stromal stem cells with pharmaceutically acceptable biodegradable materials.

[0060] The tissue-engineered cartilage formed by the method of the present invention is divided into two types, one is a compound composed of a mixture of chondrocytes and bone marrow stromal stem cells and injectable biomaterials, and the compound can be directly injected into subcutaneous and cartilage defects and other parts of the body. The other is a composite composed of a mixture of chondrocytes and bone marrow stromal stem cells and non-injectable biomaterials, which can be directly implanted into subcutaneous tissue, cartilage defects, and other parts of the body.

[0061] In a preferred embodiment the biodegradable material is solid. At this time, the above-mentioned cells and culture mediu...

Embodiment 1

[0072] Acquisition and cultivation of bone marrow stromal stem cells

[0073] Experimental animals: 5 young pigs, weighing about 10kg, and 10 adult nude mice, all provided by Chuansha Breeding Factory.

[0074] (1) Materials

[0075] 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 pentobarbital sodium (2.5% ) 1ml / kg anesthetized, routinely sterilized and draped, punctured the proximal femur with a 16-gauge needle, extracted 6-8ml of bone marrow, and placed it in a heparinized centrifuge tube.

[0076] (2) Separation of nucleated cells (density gradient centrifugation)

[0077]Repeatedly aspirate the extracted bone marrow several times with 5ml and 1ml empty needles successively, transfer to another centrifuge tube, add a small amount of serum-free DMEM culture medium, mix well, centrifuge at 3000rpm for 10 minutes, gently absorb fat and mos...

Embodiment 2

[0084] Chondrocyte isolation and culture

[0085] Anesthetize the animal as before, remove the auricle under aseptic conditions, peel off the perichondrium, cut into 5mm×5mm pieces, wash twice with phosphate buffer saline (PBS, containing 100U / ml of penicillin and streptomycin), and press the modified According to the Klagsbrun method, add 2 times the volume of 1mg / ml type II collagenase (Worthington, Freehold, NJ, USA), place it in a constant temperature shaker at 37°C for digestion for 8-12 hours, filter, and centrifuge; the precipitated cells are washed twice with PBS , make cell suspension with DMEM culture medium containing 10% fetal bovine serum, L-glutamine 300ug / ml, vitamin C 50ug / ml, penicillin and streptomycin 100U / ml, count and check with trypan blue staining Chondrocyte viability, general viability > 85%, with 2.7×10 4 cells / cm 2 Inoculate in a 10 cm diameter Petri dish at 37 °C, 5% CO 2 and 100% saturated humidity incubator for 3 days, then replace the culture ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention discloses a tissue engineering soft bone graft. It includes: (a). pharmaceutically-acceptable bio-degradable material; and (b) mixture of soft bone cell and bone marrow stemcells, BMSCs, and the mixing ratio of the described soft bone cell and bone marrow stemcells is 1:9-5:5. Said invention can effectively cure defect of soft bone. Said invention also provides its preparation method and application.

Description

technical field [0001] The invention relates to the fields of medicine and biomedical engineering, and more specifically relates to tissue engineered cartilage and its preparation method and application. Background technique [0002] As a kind of connective tissue, cartilage mainly supports, protects, and disperses stress. Histologically, it can be divided into three types: hyaline cartilage (articular cartilage, costal cartilage, nasal cartilage, tracheal cartilage), elastic cartilage (epiglottic cartilage, ear cartilage), and fibrocartilage (meniscus). Histologically, they are all single tissues containing only chondrocytes, without blood vessels inside, and their nutrition mainly depends on the penetration of interstitial fluid. [0003] Structurally, chondrocytes are wrapped in the cartilage lacuna, and the thick cartilage capsule around it acts as a natural barrier to separate chondrocytes from surrounding tissues. Therefore, cartilage tissue has weak antigenicity, is...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/00
Inventor 曹谊林周广东刘伟崔磊
Owner 上海第二医科大学附属第九人民医院
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap