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A kind of inducer of stem cell differentiation into chondrogenic and its application

A technology of differentiation induction and inducer, applied in the field of biomedicine, which can solve the problems of shortening the induction differentiation cycle to less than one week, complicated operation process, and inaccurate preparation ratio, and achieves simple and efficient induction method, simple preparation process, and shortened culture cycle Effect

Active Publication Date: 2022-04-22
BIOISLAND LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether it is to directly transplant MSCs to induce chondrogenic differentiation in situ or to construct cartilage in vitro through cartilage tissue engineering for transplantation and replacement, neither of the two ideas can avoid a key step, that is, to induce chondrogenic differentiation of MSCs.
[0005] At present, the induction of MSCs into cartilage generally uses inducers containing dexamethasone, transforming growth factor β1 (TGF-β1), ascorbic acid, sodium pyruvate, etc. and supplements other auxiliary components. A preparation for inducing stem cells to differentiate into chondrocytes by TGF-β, dexamethasone and vitamin C, CN109825469A discloses a chondrogenic differentiation inducer containing TGF-β1, ferritin, ascorbic acid, dexamethasone and sodium pyruvate, adding factors There are many types, which easily lead to inaccurate preparation ratios and increased costs, and MSCs need to be induced for 14 to 21 days before MSCs can show obvious morphological characteristics and positive staining results; Improved, the induction time was shortened to 8 days, but there are still problems such as a wide variety of added factors and complicated operation procedures
[0006] The existing technology is still unable to shorten the induction and differentiation period of MSCs to chondrogenic to less than one week

Method used

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  • A kind of inducer of stem cell differentiation into chondrogenic and its application
  • A kind of inducer of stem cell differentiation into chondrogenic and its application
  • A kind of inducer of stem cell differentiation into chondrogenic and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0070] In this example, human umbilical cord mesenchymal stem cells (hUMSCs) were used as the cell type, and DMEM / F12 was selected as the basal medium to induce chondrogenic differentiation. The steps are as follows:

[0071] (1) Collect the umbilical cords of full-term healthy cesarean section fetuses, immerse them in PBS or saline containing 1% penicillin and streptomycin, and place them on ice;

[0072] (2) Cut the umbilical cord into small sections with a length of about 3 cm on the ultra-clean table, cut them longitudinally, rinse repeatedly with sterile PBS until the liquid is free of blood, separate the arteries and veins (3 in total), and the remaining tissue is Huatong glue and amnion;

[0073] (3) Cut the remaining tissue to less than 2mm 3 After washing with PBS, place the tissue block in DMEM / F12 medium containing 15% fetal bovine serum at 37°C, 5% CO 2 Cultivated in an incubator;

[0074] (4) Observe the creeping out of the cells around the tissue block under a...

Embodiment 2

[0084] In this example, human umbilical cord mesenchymal stem cells (hUMSCs) were used as the cell type, and DMEM high glucose was selected as the basal medium to induce cartilage differentiation. The DMEM high-glucose medium of mLTGF-β1 and 10 μg / mL insulin, other conditions and experimental steps are the same as in Example 1.

[0085] Such as figure 2 As shown, after 3 days of culture, the cell morphology of hUMSCs can be significantly changed, and the cells aggregate to form a classic tissue-like pattern of chondrocyte differentiation, which can be efficiently differentiated into chondrocyte-like tissue groups, indicating that the differentiation induction medium of the present invention can efficiently induce MSCs within 3 days Differentiate into chondrocyte clusters.

Embodiment 3

[0087] In this example, human bone marrow mesenchymal stem cells (hBMSCs) were used as the cell type, and α-MEM was selected as the basal medium to induce cartilage differentiation. mL TGF-β1 and 10 μg / mL insulin α-MEM medium, other conditions and experimental steps are the same as in Example 1.

[0088] Such as image 3 As shown, after 3 days of culture, the cell morphology of hBMSCs can be significantly changed, and the cells aggregate to form a classic tissue-like pattern of cartilage differentiation, and they are efficiently differentiated into chondrocyte-like tissue groups, indicating that the induction differentiation medium of the present invention can efficiently induce MSCs within 3 days Differentiate into chondrocyte clusters.

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Abstract

The present invention provides an inducer for stem cell differentiation to chondrogenic and application thereof. The inducer includes PDGF, bFGF and TGF-β; the inducer also includes insulin and / or IGF. PDGF, bFGF, TGF-β and insulin or IGF of the present invention, under the condition of high-concentration ratio, exert the multi-factor network regulation effect, synergistically induce the differentiation of mesenchymal stem cells to chondrocytes efficiently within 3 days, the formula It is concise and has a simple preparation process, which helps to significantly shorten the production cycle of chondrocytes.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to an inducer of stem cell differentiation to chondrogenic and application thereof. Background technique [0002] The damage of articular cartilage seriously affects the quality of life of patients. Due to the limited self-repair ability, the damaged articular cartilage cannot repair itself. Clinical treatment usually adopts conservative strategies, or uses biomaterial substitutes to replace damaged articular cartilage, but there are still many problems and the goal of complete repair cannot be achieved. [0003] In recent years, the development of stem cell therapy technology has provided unlimited possibilities for the treatment of degenerative or injury diseases. Mesenchymal stem cells (MSCs) are a type of cells with self-renewal and differentiation capabilities, and more and more studies have confirmed their potential in the treatment of various diseases. There are many sour...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077A61K35/32A61P19/08
CPCC12N5/0655A61K35/32A61P19/08C12N2501/135C12N2501/115C12N2501/15C12N2501/105C12N2501/33
Inventor 徐洪杰秦大江
Owner BIOISLAND LAB
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