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Method for immortalizing human exfoliated deciduous tooth endodontium stem cell line through hTERT recombinant lentivirus

A technique for exfoliating deciduous tooth pulp and recombinant lentivirus, which can be applied to biochemical equipment and methods, botanical equipment and methods, and cells modified by introducing foreign genetic material, which can solve the loss of cell biological function and low transfection efficiency. , poor representation of cell models, etc., to achieve the effect of good function and stable properties

Inactive Publication Date: 2015-11-18
XI AN JIAOTONG UNIV
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Problems solved by technology

[0008] At present, compared with other methods to establish immortalized cells, the transfection efficiency is low, which will cause partial loss of cell biological functions and the shortcomings of transformation characteristics, and it is poorly represented as a cell model.

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  • Method for immortalizing human exfoliated deciduous tooth endodontium stem cell line through hTERT recombinant lentivirus
  • Method for immortalizing human exfoliated deciduous tooth endodontium stem cell line through hTERT recombinant lentivirus
  • Method for immortalizing human exfoliated deciduous tooth endodontium stem cell line through hTERT recombinant lentivirus

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Embodiment Construction

[0036] The following describes the specific implementation process of the present invention. It should be noted that these embodiments have only illustrative effects on the present invention, but not restrictive effects.

[0037] The method of hTERT recombinant lentivirus immortalized human shedding deciduous tooth dental pulp stem cell line includes the following steps:

[0038] Step 1. Primary culture of SHED stem cells from primary human shedding deciduous teeth:

[0039] Collect the retained deciduous incisors extracted from the Department of Pediatric Dentistry, use a pulp extraction needle to extract the pulp tissue as completely as possible in the ultra-clean table, and mix 3mg / ml type I collagenase and 4mg / ml Dispase enzyme according to the volume ratio of 1:1 Put it in a petri dish, use ophthalmological scissors to cut the pulp tissue as much as possible, and place it at 37℃, 5% CO 2 In a standard environment with saturated humidity, the cell incubator is digested for 60 mi...

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Abstract

The invention relates to a method for immortalizing a human exfoliated deciduous tooth endodontium stem cell line through hTERT recombinant lentivirus. The method comprises: 1, carrying out primary culture on primary human exfoliated deciduous tooth endodontium stem cells SHED; and 2, constructing immortalized human exfoliated deciduous tooth endodontium stem cells hTERT-SHED stably expressing hTERT gene. According to the present invention, the constructed immortalized genetically engineered cells expressing the hTERT gene is adopted as the effective cell model for researching the biological characteristics of the human exfoliated deciduous tooth endodontium stem cells so as to easily and deeply research the applications of the constructed immortalized genetically engineered cells as the seed cells in the periodontal tissue engineering and the actions of different drugs on the cells and the signal transduction pathways of the related action mechanisms, and the constructed immortalized genetically engineered cells are used to screen the traditional Chinese medicine effective components for effectively treating chronic periodontitis so as to provide the powerful tool for the subsequent research of the periodontal tissue repair and reconstruction and the development of the novel effective drugs.

Description

Technical field [0001] The invention belongs to the technical fields of cell engineering, genetic engineering and immortalization, and particularly relates to a method for hTERT recombinant lentiviral immortalization of human exfoliated deciduous teeth dental pulp stem cell lines, mainly for the primary culture and identification of human exfoliated deciduous teeth dental pulp stem cell lines, and through lentivirus The vector infects the human telomerase reverse transcriptase hTERT gene to construct an immortalized human exfoliated deciduous tooth dental pulp stem cell line, and examines its function and transformability. Background technique [0002] The harm of periodontitis has attracted more and more attention. With the development of periodontitis, it can lead to the loss of periodontal attachment, the destruction and absorption of alveolar bone, the loosening of teeth and even the loss of tooth loss. With the development of periodontal disease research, it has been found t...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N5/10
Inventor 李昂刘瑾苟建重尹战海
Owner XI AN JIAOTONG UNIV
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