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Cell culture fluid, application thereof and method for inducing periodontal ligament stem cells to differentiate into myocardium-like cells

A technology of periodontal ligament stem cells and cardiomyocyte-like cells, applied in the field of stem cell culture, can solve the problems of high medical expenses, weak value-added ability, and irreversible cardiomyocytes.

Inactive Publication Date: 2017-01-11
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although clinical drugs and interventional therapy can improve symptoms, they cannot restore damaged cardiomyocytes
Although heart transplantation can fundamentally solve the problem of ventricular remodeling, it is difficult to be widely used because of limited donors and high medical costs
Studies in recent years have shown that cardiomyocytes also have the ability to regenerate. After myocardial infarction, cardiomyocytes can also divide and proliferate. However, due to their weak proliferative ability, they cannot meet the requirements to make up for the loss of cardiomyocytes and reverse ventricular remodeling. process
There is no research report on the induction of PDLSCs differentiation into cardiomyocytes

Method used

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  • Cell culture fluid, application thereof and method for inducing periodontal ligament stem cells to differentiate into myocardium-like cells
  • Cell culture fluid, application thereof and method for inducing periodontal ligament stem cells to differentiate into myocardium-like cells
  • Cell culture fluid, application thereof and method for inducing periodontal ligament stem cells to differentiate into myocardium-like cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 PDLSCs primary isolation culture, subculture and identification

[0044] Material collection: Take the periodontal ligament tissue from the middle and lower 1 / 3 of the tooth root, transfer it into a centrifuge tube, and cut the tissue piece into about 1mm with ophthalmic scissors 3 size.

[0045] Digestion: add an appropriate amount of collagenase-dispase enzyme 1:1 mixed digestion solution (collagenase 3g / L, dispase enzyme 4g / L), and place it in a constant temperature shaker at 37°C for 30-35min. After digestion, centrifuge at 1000r / min for 5min and discard the supernatant. Add an appropriate amount of PBS to resuspend, centrifuge at 1000r / min for 5min, and discard the supernatant.

[0046] Inoculation: add culture medium (DMEM / F12 culture medium containing 10% FBS by volume) to resuspend cells, inoculate in six-well plate, 5% CO 2 Culture in an incubator at 37°C.

[0047] Purification: when the cells are 80% to 90% full, the cells are collected, and the ...

Embodiment 2

[0054] The differentiation of embodiment 2PDLSCs

[0055] The formula of each test group's culture solution is shown in Table 2:

[0056] Table 2 The formulation of each component differentiation induction solution

[0057] group basal medium FBS 5-aza Ang-II Experimental group 1 DMEM / F12 10％ 5umol / L 1umol / L Experimental group 2 DMEM / F12 10％ 10umol / L 5umol / L Experimental group 3 DMEM / F12 10％ 5umol / L 10umol / L Experimental group 4 DMEM / F12 10％ 15umol / L 10umol / L Control group 1 DMEM / F12 10％ —— —— Control group 2 DMEM / F12 10％ 10umol / L —— Control group 3 DMEM / F12 10％ —— 5umol / L

[0058] Select the 3rd generation PDLSCs prepared in Example 1, press 1×10 4 The cell / mL density was inoculated in a six-well plate with polylysine-treated sterile coverslips, and 2 ml of DMEM / F12 medium containing 10% FBS was added to each well. After 24 hours, the corresponding culture medium of each gr...

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Abstract

The invention relates to the technical field of stem cell culture and especially relates to a cell culture fluid, an application thereof and a method for inducing periodontal ligament stem cells to differentiate into myocardium-like cells. The cell culture fluid provided by the invention contains a basic culture fluid, FBS, 5-aza and Ang-II. The cell culture fluid provided by the invention can increase the differentiation rate of the periodontal ligament stem cells differentiating into myocardium-like cells and can shorten the differentiating time. A result of calculation for the differentiation rate of each group of cells shows that the differentiation rate of experimental group cells is obviously higher than that of the control group (p is less than 0.01).

Description

technical field [0001] The invention relates to the technical field of stem cell culture, in particular to a cell culture solution and its application and a method for inducing periodontal ligament stem cells to differentiate into cardiomyocyte-like cells. Background technique [0002] Myocardial infarction is the myocardial damage caused by the imbalance between coronary blood flow and myocardial demand caused by changes in coronary circulation. It is a serious clinical ischemic heart disease. Cardiomyocytes after necrosis are gradually replaced by scar tissue. Due to the lack of elasticity of scar tissue, it is difficult to meet the requirements of cardiac contraction and relaxation. In order to compensate for the loss of cardiomyocyte function, the heart gradually undergoes degenerative left ventricular remodeling. decline, eventually leading to congestive heart failure and even sudden death. Although clinical drugs and interventional therapy can improve symptoms, they c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0657C12N2501/06C12N2501/32C12N2506/1361
Inventor 陈海佳葛啸虎王一飞戚康艺张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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