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Kit and method for identifying periodontal ligament stem cells

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: 2021-03-16
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 periodontal ligament stem cells
  • Kit and method for identifying periodontal ligament stem cells
  • Kit and method for identifying periodontal ligament stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0074] 1. This embodiment employs dental stem cell samples from 3 groups of different individuals, and corresponding and pulp stem cells samples.

[0075] Cultured periodontal stem cells and pulp stem cells, and extracting a full protein after cell cleavage. DTT, IAA, and pancreatic enzyme were then added, and the peptide segment was obtained after 37 ° C overnight. The TMT tag was then added, labeled at normal temperature, and the reaction was reflected after 15 min after 5% hydroxylamine. After drying, the desalination was dried again, and the detection of the peptide segment was performed on LC-MS / MS.

[0076] 2, experimental results

[0077] The statistical results show that the CKB protein in the periodontal permeable stem cell group is higher, the pulp stem cell group CKB is low (eg figure 1 Indicated).

Embodiment 2

[0078] Example 2 Identification of periodontal permeable stem cells and pulp stem cell identification markers

[0079] 1. Total RNA extraction of periodontal stem cells and pulp stem cells

[0080] This test uses a dental permeable stem cell sample from 3 groups of different individuals, and the corresponding pulp stem cell samples.

[0081] 1) Pour the culture medium in the culture dish / bottle, PBS wash 2-3 times, add 0.7 ml Trizol solution (red), bleate the cells attached to petri dish / bottle with a gun, transfer to 1.5 mL centrifugation In the tube, the petri dish was added to 0.5 mL of Trizol flushing petri dish harvested the remaining cells that were not fully blown, transferred to the same centrifugal tube to stand for 5 min.

[0082] 2) Direct addition of 200 μL of chloroform, vortex oscillating 15 s mixed back room temperature 2-3 min (or add chloroform to make the protein, DNA, and lipids).

[0083] 3) 4 ° C, 12000 rpm centrifuge for 20 min.

[0084] 4) After centrifuga...

Embodiment 3

[0098] Example 3 Identification of CKB markers in periodontal permeable stem cells and gum stem cells

[0099] The method employed in this example is not different from Example 2, and the identification of CKB markers by dental stem cell samples from 3 groups of different individuals, respectively, using a dental stem cell sample, respectively.

[0100] result:

[0101] The average value of the periodontal membrane stem cell sample group and the average value of the pulp stem cell sample group were compared. MRNA expression test results show that the expression of CKB in the periodontal permeable stem cell group is high, and the CKB of gums in the gums is low (eg image 3 Indicated).

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Abstract

The present invention provides a kit and method for identifying periodontal ligament stem cells, which relate to the field of biotechnology. The inventors of the present invention have found through experiments that CKB gene and / or CKB protein have a characteristic of significantly high expression in periodontal ligament stem cells Therefore, periodontal ligament stem cells can be accurately and effectively identified by detecting the expression of CKB gene and / or CKB protein in the cell sample to be tested.

Description

Technical field [0001] The present invention relates to the field of biotechnology, and more particularly to a kit and method for identifying a periodontal stem cell. Background technique [0002] With the rise of stem cells and regenerative medicine, the basis and application of stem cells have emerged in endless. From embryonic stem cells, hematopoietic stem cells, bone marrow stem cells to induce multifunctional stem cells, are verified as potential clinical materials. Oral stem cells have started to rise in recent years. Starting in the 21st century, there were 8 stem cells from human dental organizations from human dentistry. [0003] Periocelerated stem cells (PDLSCs) can be obtained from adult dental membrane tissues, and materials can be derived from dental surgical discards. Studies have shown that PDLSCs can also have high proliferation and multi-directional potential compared to other stem cells, and can differentiate into bone, sugne, and the like under appropriate co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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