Method for inducing directed cartilage differentiation of stem cells
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A technology of stem cells and cartilage, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve problems such as decomposition, high price, and influence on characteristics, achieve good cartilage differentiation ability, prevent protease decomposition, and help effect on secretion
Active Publication Date: 2020-01-14
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
View PDF3 Cites 1 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
At present, the commonly used methods for delivering growth factors to effectively induce chondrogenic differentiation of stem cells are: 1) Add protein growth factors exogenously, but protein growth factors are expensive and unstable in the body, easily decomposed by proteases, lack of long-term stability
2) Through the design and preparation of biological materials, growth factors are loaded and controlled release, but the loading process and conditions will have a certain impact on the characteristics of macromolecular growth factors
However, although non-viral vectors can be operated safely, the transfection efficiency has not been ideal; while viral vectors have high transfection efficiency, but the transfection process will cause some damage to the seed cells, and their safety is still controversial.
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
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0033] Example 1 Transfection of adenovirus carrying TGF-β3 into chondrocytes
[0035] Purchase the Adeno-XTM ViraTrakTM ZsGreen1 Expression System 2 kit, add Xhol / NotI recognition sites at both ends of the transforming growth factor TGF-β3 sequence by PCR, and then use DNA recombination technology to insert the TGF-β3 sequence into the adenovirus vector ( Green fluorescent protein adenoviral vector), construct an adenoviral vector (Ad-T3) carrying TGF-β3, Ad-T3 amplified to >10 9IFU / mL and then frozen for later use. The specific construction method refers to the following literature: Hao, J. H., Yao, Y. C., Varshney, R. R., Wang, L. C., Prakash, C., Li, H., Wang, D. A., GeneTransfer and Living Release of Transforming Growth Factor-Beta 3 for Cartilage Tissue Engineering Applications, Tissue Engineering Part C Methods, 14 (2008), 4: 273-280.
[0036] (2) Isolation and culture of chondroc...
Embodiment 2
[0039] Example 2 Observation of the culture process of ATDC5 cells and transfected chondrocyte co-culture system (named 3A1C) and detection of directional cartilage differentiation
[0040] (1) Purchase ATDC5 cells from Abgent (San Diego) (this cell is recognized as a substitute for stem cells as model cells), using F12 medium [containing 0.5% FBS (fetal bovine serum) and 1% penicillin-streptomycin penicillin-streptomycin (Double Antibody)] Passage ATDC5 cells to 3 passages and freeze them for later use;
[0041] (2) ATDC5 cells and transfected chondrocytes were digested, collected, counted, mixed at a ratio of 3:1, and 1.2% sodium alginate solution was added to prepare a cell density of 6×10 6 cells / mL of cell suspension. Take 50uL of the above cell suspension with a special pipette tip, and quickly inject it into 102mM CaCl 2 solution to form a cell gel (results image 3 ), and after 10 min, remove the cell gel with a long spoon and place it on a 24-well plate, one capsul...
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
Property
Measurement
Unit
quality score
aaaaa
aaaaa
Login to view more
Abstract
The invention belongs to the technical field of tissue engineering, and particularly relates to a method for inducing directed cartilage differentiation of stem cells. Based on a co-culture system ofcartilage cells and the stem cells, the cartilage cells and the stem cells are subjected to division and cooperation of labor through the co-culture system, an adenovirus vector is adopted to introduce a transforming growth factor (TGF-beta 3) into the stem cells, the continuous, local and over-expressed TGF-beta 3 are provided for the stem cells through transfected cartilage cells, directed cartilage cell differentiation of the stem cells is stimulated, exogenous addition of TGF-beta 3 is not needed, thus the growth factor is prevented from being decomposed by protease, damage possibly generated in the gene transfection process to the stem cells is avoided, and the higher survival rate and the better cartilage differentiation ability of the stem cells are maintained. In addition, throughthe mode of co-culture of the cartilage cells and the stem cells, directed cartilage differentiation of the stem cells in a cartilage cell conditional culture medium is facilitated, induction to the growth factor is acted more sensitively, and secretion of cartilage epimatrix is more facilitated.
Description
technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a method for inducing directional cartilage differentiation of stem cells. Background technique [0002] Articular cartilage has the functions of buffering mechanical impact, reducing friction between joints, and increasing joint adaptability. Its slight damage will lead to osteoarthritis (OA). Since articular cartilage is a tissue without nerves and blood vessels, its self-repair ability very bad. At present, there are many methods for repairing cartilage damage, but each has its own defects, and none of them can solve this problem well. Moreover, chondrocytes are surrounded by extracellular matrix, so it is difficult for exogenous drugs or various factors to act on it. Therefore, the repair of articular cartilage damage is still a medical problem. [0003] In recent years, with the rapid development of tissue engineering, some therapeutic strategies ha...
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
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more