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Application of LncRNA-TUG1 in preparation of drug for regulating and controlling dryness maintenance capacity of periodontal ligament stem cells

A technology of periodontal ligament stem cells and ability, applied in the application field of medicine, can solve the problems of reporting the stemness maintenance ability of periodontal ligament stem cells, and achieve the effect of precision medicine

Active Publication Date: 2017-11-07
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there is no report about LncRNA regulating the stemness maintenance ability of human periodontal ligament stem cells

Method used

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  • Application of LncRNA-TUG1 in preparation of drug for regulating and controlling dryness maintenance capacity of periodontal ligament stem cells
  • Application of LncRNA-TUG1 in preparation of drug for regulating and controlling dryness maintenance capacity of periodontal ligament stem cells
  • Application of LncRNA-TUG1 in preparation of drug for regulating and controlling dryness maintenance capacity of periodontal ligament stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Research on the biological characteristics of periodontal ligament stem cells

[0036] 1. Isolation and culture of human periodontal ligament stem cells

[0037] Under anesthesia, the human impacted third molar was aseptically extracted, and the periodontal tissue around it was gently peeled off. The periodontal tissue in the middle was taken, washed repeatedly with PBS, cut into small pieces, and placed in a solution containing type Ⅰ collagenase (3g / L) and Dispase II enzyme (4g / L) digestion solution, digested at 37°C for 40 minutes, collected cells through a 70um cell sieve, centrifuged at 1000 rpm for 5 minutes, removed the upper liquid, and resuspended into a single cell suspension with fresh culture medium liquid. Cells were seeded at 25cm 2 In a cell culture flask, in α-MEM medium (containing 15% fetal bovine serum, 2 mmol / L glutamine, 100 U / ml penicillin and 100 μg / ml streptomycin) at 37 °C, 5% CO 2 For culture, the medium was changed every 2-3 days...

Embodiment 2

[0046] Example 2: Regulatory effect of LncRNA-TUG1 on the stemness maintenance ability of periodontal ligament stem cells

[0047] 1. Construction of lentiviral vector:

[0048] Primers were designed according to the human LncRNA-TUG1 sequence (ID: 55000), and the primers were synthesized and purified by Shanghai Lingke Biotechnology Co., Ltd. After annealing, it was ligated to the linearized pGenesil-1 plasmid to construct the lncRNA-TUG1 (silencing) expression vector lentivirus to inhibit the expression of LncRNA-TUG1.

[0049] details as follows:

[0050] Vector: pLenO-GFP

[0051] Lenti-human-NC sequence: TTCTCCGAACGTGTCACGT (SEQ ID NO: 1); (NegativeControl).

[0052] Lenti-human-sh1 TUG1

[0053] GPH-hTUG1-SH1 Forward Sequence

[0054] 5'-GATCCCTGTTGACCTTGCTGTGAGAACTTCCTGTCAGATTCTCACAGCAAGGTCCAACAGTTTTTG-3' (SEQ ID NO: 2);

[0055] GPH-hTUG1-SH1 Reverse Sequence

[0056] 5'-AATTCAAAAACTGTTGACCTTGCTGTGAGAATCTGACAGGAAGTTTCCACAAGGTCAACAGG-3' (SEQ ID NO: 3).

[0057] ...

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PUM

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Abstract

The invention discloses application of LncRNA-TUG1 in the preparation of a drug for regulating and controlling the dryness maintenance capacity of periodontal ligament stem cells. The invention discovers that LncRNA-TUG1 play very important regulation and control roles in the dryness maintenance process of the periodontal ligament stem cells for the first time. When LncRNA-TUG1 is decreased, the proliferation, self-renewal, differentiation and transfer capacities of PDLSCs are inhibited, so that LncRNA-TUG1 is determined to be taken as a potential target for the dryness maintenance of multi-energy stem cells, the development of new drugs or factors relevant with is promoted, LncRNA-TUG1 is finally applied to field of regenerative medicines, and the true accurate medical treatment on clinic is achieved.

Description

technical field [0001] The invention relates to the technical field of oral tissue regeneration, in particular to the application of LncRNA-TUG1 in the preparation of medicines for regulating the stemness maintenance ability of periodontal ligament stem cells. Background technique [0002] Periodontal tissue regeneration is one of the key points of stomatology research, and it is the common goal of periodontics, maxillofacial surgery, orthodontics, prosthodontics and other multidisciplinary research fields. In recent years, the research progress on dental stem cells and tissue engineering technology has brought new research directions to the biological regeneration of periodontal tissue. In 2004, Seo et al. used single cell cloning technology to successfully isolate and identify a new type of adult stem cells from human periodontal ligament tissue, that is, periodontal ligament stem cells, which opened up new ideas for the restoration of oral biological functions. [0003] ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61P1/02C12N15/867
CPCA61K45/00C12N15/86C12N2740/15043
Inventor 魏福兰何琴马晓妮李梦莹
Owner SHANDONG UNIV
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