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
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[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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