Cartilage cell treating agent comprising collagen, hyaluronic acid derivative, and stem cell derived from mammal umbilical cord

A chondrocyte and hyaluronic acid technology, applied in the field of chondrocyte therapeutics, can solve problems such as cell leakage, preventing tissue regeneration, and inability to adequately treat diseased areas.

Inactive Publication Date: 2014-05-14
车比奥及戴奥斯泰有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, this method has limitations because when the damaged area is large, cultured cells alone may not be able to adequately treat the affected area and may be squeezed by weight allowing cells to leak out preventing tissue regeneration

Method used

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  • Cartilage cell treating agent comprising collagen, hyaluronic acid derivative, and stem cell derived from mammal umbilical cord
  • Cartilage cell treating agent comprising collagen, hyaluronic acid derivative, and stem cell derived from mammal umbilical cord
  • Cartilage cell treating agent comprising collagen, hyaluronic acid derivative, and stem cell derived from mammal umbilical cord

Examples

Experimental program
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Embodiment 1

[0072] Analysis of Separation and Proliferation Efficiency of Umbilical Cord Stem Cells

[0073] The umbilical cords used in this study were discarded after delivery and obtained with the consent of normal mothers. Use the umbilical cord within 24 hours of collecting it.

[0074] Tissues from the blood outside the umbilical cord were removed by using Ca-free 2+ and Mg 2+ DPBS to remove the outer amniotic membrane and two arteries, cut to 1mm 3 size, and then placed in α-minimal essential medium (α-MEM) containing 100 U / mL penicillin, 0.1 μg / mL streptomycin and 0.2 μg / mL umbilical cord extract. After culturing them for 7 days, when the cells appeared attached to the bottom, the cells were treated with α-MEM containing 500 U / ml type I collagenase for 4 hours to detach the cells. Afterwards, the cells were divided into 2 × 10 3 piece / cm 2 Inoculate the culture dish (containing α-MEM, the dish contains 100U / ml penicillin, 0.1μg / ml streptomycin and 0.2mg / ml umbilical cord ex...

Embodiment 2

[0075] Analysis of embryonic stem cell markers by RT-PCR

[0076] Cell pellets were treated with Ca-free 2+ and Mg 2+ was washed with DPBS, 1 ml of lysis buffer (iNtRON Biotechnology product) was added thereto, and then total RNA was isolated therefrom according to the method described in the manual available from iNtRON Biotechnology. 1 μg of RNA was reverse-transcribed in 20 μL of reaction solution containing reaction buffer, 1 mM dNTP mixture, 0.5 μg / μL oligo(dT)15, 20 U RNase inhibitor and 20U of AMV reverse transcriptase. The reaction was carried out at a temperature of 42°C for 60 minutes. The RT product (cDNA) obtained therefrom was subjected to PCR using a 2X PCR Master mixed solution kit (product of iNtRON Biotechnology), which contained 10 μL of a reaction solution containing 1X Taq buffer, 0.25 U Taq polymerase, 10 pM Sense and antisense gene-specific primers. Amplification was performed for a total of 32 cycles and each cycle included denaturation at 94°C for...

Embodiment 3

[0081] Analysis of the expression of mesenchymal stem cell markers by FACS

[0082] Flow cytometry was used to analyze the characteristics of the isolated cells. The isolated cells were washed with PBS, treated with trypsin-EDTA to prepare single-cell groups, and then washed with PBS containing 2% FBS and 1 mM EDTA. Thereafter, stem cell markers bound to fluorescein isothiocyanate (FITC) or phycoerythrin (PE) were treated and allowed to react for 20 minutes, and then analyzed with FACSCalibur (product of Becton-Dickinson).

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Abstract

The present invention relates to a medical composite biomaterial. More specifically, the present invention relates to the medical composite biomaterial comprising collagen and a hyaluronic acid derivative. Also, the present invention relates to a cartilage cell treating agent using the biomaterial and stem cells derived from a mammal umbilical cord. The biomaterial does not cause an immune system reaction, has superior durability, and the cartilage cell treating agent comprising the biomaterial and the stem cells enables arthroscopic surgery, thereby reducing pain of the patient, and can effectively treat degenerative arthritis and cartilage damage.

Description

[0001] related application [0002] This application claims the benefit of Korean Patent Application No. 10-2011-0069551 filed on Jul. 13, 2011 at the Korean Intellectual Property Office, the disclosure of which is incorporated herein by reference in its entirety. field of invention [0003] One or more embodiments of the present invention relate to a biomaterial comprising collagen and hyaluronic acid derivatives, and a chondrocyte therapy comprising the biomaterial and umbilical cord-derived stem cells. Background of the invention [0004] Collagen is the most ubiquitous protein found in humans, and the most abundant protein in mammals, constituting about 25% to about 35% of all proteins in the body. More specifically, collagen is an important component of bones, tendons, and ligaments, and mainly maintains the structure of organs. Collagen can be easily extracted from bovine or porcine skin. However, since collagen is derived from non-human animals, collagen has problem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/44A61K35/12A61K31/728A61P19/00A61K35/51A61K35/545
CPCA61K31/728A61K35/51A61K35/545A61K38/39A61L27/26A61L27/3834A61L27/3852A61L27/52A61L2400/06A61L2430/06A61P19/00A61P19/08A61P43/00A61K2300/00C08L89/06C08L5/08A61K35/12A61K35/44A61K47/36
Inventor 金仙美李荣濬崔容秀
Owner 车比奥及戴奥斯泰有限公司
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