Method for screening new subset of periodontal ligament specific stem cells based on single cell sequencing and application thereof

A single-cell sequencing and periodontal ligament technology, applied in the biological field, can solve the problems that periodontal tissue regeneration and repair cannot be improved

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A single-cell sequencing and periodontal ligament technology, applied in the biological field, can solve the problems that periodontal tissue regeneration and repair cannot be improved

CN112481364AActive Publication Date: 2021-03-12SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE

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Embodiment 1

[0025] The present invention will be further described below in conjunction with specific examples, but the examples are only exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solutions of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the protection scope of the present invention. Example 1 Method for screening new subgroups of periodontal ligament-specific stem cells based on single-cell sequencing

[0026] The present invention provides a method for screening new subpopulations of periodontal ligament-specific stem cells based on single-cell sequencing, comprising the following steps:

[0027] a. Select healthy premolars that need to be extracted due to orthodontics. The teeth are required to be complete in shape, free from c...

Embodiment 2

[0063] Embodiment 2 Experimental result analysis

[0064] (1) Single-cell transcriptome sequencing analysis revealed new subpopulations

[0065] According to the analysis of single-cell transcriptome sequencing, the tissue map of the periodontal ligament was drawn and divided into 14 subgroups, namely fibroblasts (0-4), osteoblasts (5), and new population (6). , endothelial cells (7), pericytes (8), T cells (9), epithelial cells (10), macrophages (11), neutrophils (12), monocytes (13) and possibly Schwann cells (14). From figure 2 It can be seen that single-cell transcriptome sequencing revealed that there may be a new subgroup specific to periodontal ligament tissue (population 6), figure 2 The 2D-tSNE map of A shows the coloring results of single-cell RNA-seq clustering analysis of 5562 periodontal ligament cells, and the new subpopulation of periodontal ligament tissue is circled in dark blue; the single gene tSNE map and Violin map of 2B show Known periodontal ligame...

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Abstract

The invention discloses a method for screening a new subset of periodontal ligament specific stem cells based on single cell sequencing and application thereof. The screened new subset specifically expresses FBLN2 and membrane protein CADM3, and meanwhile, the invention discloses application of the new subset of periodontal ligament specific stem cells in preparation of drugs for treating periodontal diseases. According to the invention, the subset of periodontal ligament specific stem cells is obtained through single cell sequencing, and in vitro flow sorting and cell subset performance verification are carried out through marker genes; According to the invention, the screened cell subset is beneficial to search for intervention targets of periodontal steady-state imbalance, and creates possibility for development of new drugs; periodontal ligament specific stem cells are used as seed cells to form organoids, and a scientific basis is provided for research and verification of new medical instrument materials; and the autologous / allogeneic transplantation feasibility of the periodontal ligament stem cells is further explored, and a new remodeling thought is provided for treating periodontal diseases.

Description

technical field [0001] The invention belongs to the biological field, and in particular relates to a method for screening a new subset of periodontal ligament-specific stem cells based on single-cell sequencing and an application thereof. Background technique [0002] In the complex oral microenvironment, periodontal tissue is highly dynamic. Periodontal homeostasis is the biological basis for normal oral physiological functions such as mastication and pronunciation, as well as clinical treatment methods such as orthodontics and implants. The imbalance of periodontal homeostasis can lead to loss of periodontal attachment, alveolar bone destruction, and periodontal disease. Periodontal disease has the characteristics of high incidence, easy damage, and difficult recovery. It is the most common cause of tooth loss in humans. It is closely related to a variety of systemic diseases. It has been listed by the World Health Organization as a disease that endangers human health and...

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Application Information

Patent Timeline

12 Mar 2021

Publication

CN112481364A

IPC: C12Q1/6869; C12Q1/04; A61K35/32; A61P1/02

CPC: C12Q1/6869; A61K35/32; A61P1/02; C12Q2521/107; C12Q2535/122

Inventors: 江凌勇; 代庆刚