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Application of small molecule compound 4-aminobiphenyl (4-ABP) in promoting stem cell proliferation and cartilage differentiation

An aminobiphenyl and stem cell technology, applied in the field of cell medicine, can solve problems such as no relevant reports on the effect

Pending Publication Date: 2020-02-04
深圳丹伦基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect of 4-ABP of the present invention on promoting the chondrogenic differentiation of mesenchymal stem cells has not been reported.

Method used

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  • Application of small molecule compound 4-aminobiphenyl (4-ABP) in promoting stem cell proliferation and cartilage differentiation
  • Application of small molecule compound 4-aminobiphenyl (4-ABP) in promoting stem cell proliferation and cartilage differentiation
  • Application of small molecule compound 4-aminobiphenyl (4-ABP) in promoting stem cell proliferation and cartilage differentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Effect of 4-ABP on BM-MSC proliferation.

[0041] (1) Resuscitated bone marrow mesenchymal stem cells, inoculated at 175cm 2 Add 20 mL of complete medium to the culture bottle, place it in a 37°C, 5% CO2 incubator and incubate, changing the medium every three days. Cells in the logarithmic growth phase were inoculated in multiple 24-well culture plates, and complete medium was added, and the medium was changed every three days. Group the cells from the first day of planting, divide them into 4-ABP group, KGN group and PA group, add 10µM 4-ABP, KGN and PA respectively, change the medium every three days, and keep the same chemical small molecule after changing the medium concentration.

[0042] (2) At different time points, the cells were digested with trypsin and counted with a cell technology instrument.

Embodiment 2

[0044] Effect of 4-ABP on the expression of chondrogenic-related genes in BM-MSCs.

[0045] (1) Resuscitate bone marrow mesenchymal stem cells, inoculate in a 175cm2 culture flask, add 20mL of complete medium, incubate in a 37°C, 5% CO2 incubator, and change the medium every three days. Cells in the logarithmic growth phase were inoculated in multiple 24-well culture plates, and complete medium was added, and the medium was changed every three days. After the cell density was greater than 90%, the cells were divided into 4-ABP group, KGN group and PA group.

[0046] (2) Add chondrogenic induction medium, and at the same time add 10µM 4-ABP, KGN and PA respectively, and change the medium every three days. After 14 days of induction, RNA was extracted to detect the expression of cartilage-related genes Collagen Ⅱ, Aggrecan and Lubricin.

Embodiment 3

[0048] Effects of 4-ABP on the expression of chondrogenic effect-related genes and proteins in hUCMSCs.

[0049] (1) Resuscitated human umbilical cord mesenchymal stem cells, inoculated at 175cm 2 Add 20mL of complete medium to the culture flask, place at 37°C, 5% CO 2 Incubate in the incubator and change the medium every three days. Cells in the logarithmic growth phase were inoculated in multiple 24-well culture plates, and complete medium was added, and the medium was changed every three days. After the cell density was greater than 90%, the group was divided into Vehicle group (maintenance medium only), KGN group (maintenance medium + 10µM KGN), 4-ABP group (maintenance medium + 10µM 4-ABP), PA group (Maintenance Medium + 10 µM PA).

[0050] (2) Continue to use the complete medium as the maintenance medium to continue the culture, and add 10 µM 4-ABP, KGN and PA respectively, and change the medium every three days. After 20 days of induction, RNA was extracted to detec...

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Abstract

The invention discloses application of small molecule compound 4-aminobiphenyl (4-ABP) in promoting stem cell proliferation and cartilage differentiation. Experiments prove that the 4-ABP can promoteproliferation of mesenchymal stem cells (including bone marrow, fat, umbilical cords, placenta, joint tissue and joint fluid as well as mesenchymal stem cells derived from tissue cells such as inducedpluripotent stem cells and cartilage differentiation, and can increase the gene expression levels of Collagen II, Aggrecan and Lubricin of stem cells and expression content of corresponding protein.Therefore, the invention proves that the 4-ABP has functions of promoting proliferation of mesenchymal stem cells and cartilage differentiation and is expected to become a regeneration and repair stimulation preparation or auxiliary preparation for stem cell medicated cartilage injury and diseases. The 4-ABP can be applied to MSC preparation related to cartilage regeneration and tissue engineeringand induced differentiation at the early stage of transplantation as a small molecule preparation, and lays a foundation for developing novel technical schemes and technical measures related to prevention and tretament of cartilage tissue injury and diseases.

Description

technical field [0001] The present invention belongs to the field of cell medicine, specifically, the present invention relates to 4-Aminobiphenyl (4-Aminodiphenyl, 4-ABP) to promote the proliferation and chondrogenic differentiation of various types of mesenchymal stem cells and its application in the prevention and treatment of related diseases . Background technique [0002] Stem cells and regenerative medicine are one of the most promising branches in today's medical field, providing new opportunities for hundreds of diseases that currently lack effective treatment techniques, and have huge clinical applications and market prospects. Mesenchymal stem cells (MSC) can be isolated from more than ten kinds of adult tissues such as bone marrow, fat, joint fluid, dental pulp, cartilage, or fetal tissues (amniotic membrane, umbilical cord, placenta, etc.), and are currently the most clinically accepted and effective The type of stem cells with the best safety and the most adequ...

Claims

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Application Information

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IPC IPC(8): C12N5/077A61K31/136A61P19/08A61P19/02A61P19/00
CPCC12N5/0655A61K31/136A61P19/08A61P19/02A61P19/00C12N2501/999C12N2500/38C12N2500/30C12N2501/39C12N2501/15C12N2506/1353C12N2506/1369
Inventor 张帅周海周光前李真莫翠萍
Owner 深圳丹伦基因科技有限公司
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