Isolation and culture of dental pulp stem cells from human deciduous teeth by modified enzyme digestion

doi:10.3969/j.issn.2095-4344.2013.10.014

Abstract

Abstract:

BACKGROUND: Dental pulp stem cells from human deciduous teeth greatly proliferate and cell number required by tissue engineering can be easily acquired within a short time period.
OBJECTIVE: To compare the cell harvest success rate and biological characteristics of dental pulp stem cells from human deciduous teeth between conventional enzyme digestion and modified enzyme digestion.
METHODS: Twelve caries-free deciduous incisors were selected in this study. Dental pulp stem cells were harvested from these human deciduous teen by conventional enzyme digestion method (n=6 incisors) and modified enzyme digestion method (n=6 incisors). Cell expansion days, total cell harvest, growth curves, cell doubling time, cell colony formation rate, cell surface specific markers STRO-1, CD146, CD34 and CD45 were determined among different generations of cells. Osteogenic and adipogenic differentiation of cells were determined by detecting alkaline phosphatase and dentin sialoprotein expression levels.
RESULTS AND CONCLUSION: There were significant differences in growth curves, cell expansion days, total cell harvest and cell doubling time between conventional enzyme digestion and modified enzyme digestion (P <0.05). Alkaline phosphatase and dentin sialoprotein expression levels were significantly higher in dental pulp stem cells cultured by modified enzyme digestion than by conventional enzyme digestion method. However, there were no significant differences in cell colony formation rate, cell surface specific marker expression and induced cell differentiation between modified enzyme digestion and conventional enzyme digestion (P > 0.05). Compared to conventional enzyme digestion, modified enzyme digestion can assist in in vitro expansion and culture of dental pulp stem cells from homologous deciduous teeth within a short time period and provide high-quality seed cells for tooth regeneration and is a reliable method for in vitro culture of dental pulp stem cells from deciduous teeth.

Key words: stem cells, stem cell culture and differentiation, deciduous tooth dental pulp cells, modified enzyme digestion, dentin sialoprotein, in vitro culture, cell identification, odontogenic differentiation, provincial grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Bielikezi·kadeer, Liu Yi-shan, Wang Xuan, Li Bo-qi, Ma Yan, Bi Xiao-juan, Nusilaiti·halike. Isolation and culture of dental pulp stem cells from human deciduous teeth by modified enzyme digestion[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(10): 1793-1800.

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