Technology for tooth regeneration using keratin stem cell and dental pulp stem cell

A dental pulp stem cell technology, applied in the field of organ regeneration, can solve the problems of limiting the application of tooth regeneration technology, limited sources of dental epithelium and dental mesenchyme, and achieve the effect of convenient promotion and use and expansion of sources

Inactive Publication Date: 2005-03-02
FUJIAN NORMAL UNIV
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Problems solved by technology

Although the use of dental epithelium and dental mesenchyme can be used to regenerate teeth, the source of dental

Method used

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Experimental program
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Effect test

Embodiment

[0022] 1. Isolation and in vitro culture of dental pulp stem cells

[0023] Take human molars that have been extracted in dentistry, cut the teeth under aseptic conditions, expose the pulp cavity, and take out the pulp tissue with a straw. Digest for 50 min at 37°C in digestive solution (containing 3 mg / ml type I collagenase and 4 mg / ml neutral protease). The digested pulp tissue was blown gently with a pipette into a single cell suspension, and placed in a 10cm cell culture dish for culture. The composition of the culture medium is α-MEM containing 15% fetal bovine serum, 20 mM L-glutamine, 0.1 mM vitamin C, 100 ug / ml streptomycin, and 100 units / ml penicillin. at 37°C 5% CO 2 After culturing in an incubator for 24 h, the cells were washed three times with PBS (10 mM phosphate buffer, 2.7 mM KCl 137 mM NaCl pH7.4) to remove unattached cells. The adherent growing cells were continued to be cultured, and the culture medium was changed every three days, and after 14-16 days of...

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Abstract

The present invention belongs to the field of organ regenerating technology, and is especially tooth regenerating technology with keratin stem cell and dental pulp stem cell. The technological scheme of the present invention includes: taking out dental pulp tissue form cheek tooth, digesting with digestive enzyme and transferring to culture liquid; culturing for 24 hr, thrice washing with PBS to eliminate un-adherent cell and transferring to culture liquid; further subsequent culturing the adherent cell for 14-16 days; inducing dental pulp stem cell with growth factor MBP4 and FGF10 for 24 hr, recombining in vitro cultured keratin stem cell and dental pulp stem cell to form recombinant ball; culturing the recombinant ball in culture box at 37 deg.c and with CO2 in 5 %, transplanting into oral cavity and culturing for 10-12 weeks to form tooth. The present invention has expanded material source.

Description

technical field [0001] The invention belongs to organ regeneration technology, in particular, the invention relates to the technology of utilizing keratinocyte stem cells and dental pulp stem cells to carry out tooth regeneration. Background technique [0002] In recent years, organ engineering and tissue engineering are emerging disciplines that have developed rapidly in the field of biotechnology. Its development will make medical practices such as human genetic defects, tissue repair and organ regeneration impossible under conventional medical technology a reality. Teeth are relatively simple organs in terms of their organizational structure and development and differentiation process. Compared with other organs, the development of teeth has distinctive characteristics, which is reflected in the fact that teeth are the only organs in the human body that can develop again in adults. From the perspective of organ development, the regeneration of teeth is easier and more l...

Claims

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Application Information

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IPC IPC(8): A61F2/02A61K6/02C12N5/074
Inventor 张彦定陈一平黄义德
Owner FUJIAN NORMAL UNIV
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