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Kit and method for identifying stem cells of periodontal membrane

A periodontal ligament stem cell and kit technology, which is applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc., to achieve the effect of being suitable for popularization and application and having a wide range of clinical applications

Active Publication Date: 2021-01-15
KYBIOSTEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, since a variety of stem cells can be isolated from oral tissues, they are mostly similar in immunophenotype, proliferation and differentiation capabilities, and there are no corresponding products and methods reported on the market to distinguish and characterize periodontal ligament stem cells from other cells. make a distinction

Method used

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  • Kit and method for identifying stem cells of periodontal membrane
  • Kit and method for identifying stem cells of periodontal membrane
  • Kit and method for identifying stem cells of periodontal membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Screening of periodontal ligament stem cells and dental pulp stem cell markers

[0074] 1. In this example, samples of periodontal ligament stem cells and corresponding samples of dental pulp stem cells from 3 different individuals were used.

[0075] Periodontal ligament stem cells and dental pulp stem cells were cultured, and whole protein was extracted after cell lysis. Then DTT, IAA and trypsin were added respectively, and the peptides were obtained after overnight reaction at 37°C. Then add the TMT tag, label at room temperature, add 5% hydroxylamine and react for 15 min to terminate the reaction. After desalting after drying, the peptides were detected on LC-MS / MS after drying again.

[0076] 2. Experimental results

[0077] The statistical results showed that the abundance of CKB protein in the periodontal ligament stem cell group was higher, and the abundance of CKB in the dental pulp stem cell group was lower (such as figure 1 shown).

Embodiment 2

[0078] Example 2 Identification of identification markers for periodontal ligament stem cells and dental pulp stem cells

[0079] 1. Extraction of total RNA from periodontal ligament stem cells and dental pulp stem cells

[0080] This experiment used periodontal ligament stem cell samples and corresponding dental pulp stem cell samples from 3 different individuals.

[0081] 1) Pour off the medium in the culture dish / bottle, wash with PBS 2-3 times, add 0.7 mL Trizol solution (red), blow off the cells attached to the culture dish / bottle with a gun, and transfer directly to a 1.5 mL centrifuge Add 0.5 mL of Trizol to the petri dish to wash the petri dish to harvest the remaining cells that were not completely blown off, transfer to the same centrifuge tube for shaking, and let stand at room temperature for 5 min.

[0082] 2) Add 200 μL of chloroform directly, vortex for 15 s to mix well, then place at room temperature for 2-3 min (or add chloroform to fully separate proteins, DNA...

Embodiment 3

[0098] Example 3 Identification of CKB markers in periodontal ligament stem cells and gingival stem cells

[0099] The method used in this example is not different from that in Example 2. The samples of periodontal ligament stem cells and gingival stem cells from three different groups of individuals were used to identify CKB markers.

[0100] result:

[0101] The average value of the periodontal ligament stem cell sample group was compared with the average value of the dental pulp stem cell sample group. The results of mRNA expression test showed that the expression of CKB in the periodontal ligament stem cell group was high, and the expression of CKB in the gingival stem cell group was low (such as image 3 shown).

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Abstract

The invention provides a kit and method for identifying stem cells of a periodontal membrane and relates to the technical field of biology. Discovered by an inventor of the invention through experiments, a CKB gene and / or CKB protein has the characteristic of remarkable high expression in the stem cells of the periodontal membrane, so that the stem cells of the periodontal membrane can be accurately and effectively identified through detecting expression conditions of the CKB gene and / or CKB protein in cell samples to be detected.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit and method for identifying periodontal ligament stem cells. Background technique [0002] With the rise of stem cells and regenerative medicine, basic and applied research on stem cells emerges in an endless stream. From embryonic stem cells, hematopoietic stem cells, bone marrow stem cells to induced pluripotent stem cells, all have been verified as potential clinical materials. Oral stem cells have been on the rise in recent years. Since the beginning of the 21st century, eight types of stem cells have been isolated from human dental tissues one after another. [0003] Periodontal ligament stem cells (PDLSCs), as a kind of oral stem cells, can be obtained from adult periodontal ligament tissue, and the materials can be obtained from dental surgery discards. Studies have shown that compared with other stem cells, PDLSCs also have high proliferation and multi-directional dif...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/04C12N15/11
CPCC12Q1/6888C12Q2600/158C12Q2600/166
Inventor 雷童王建张晓霜刘彦彦王治世杜宏武
Owner KYBIOSTEM CO LTD
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