FGF8/SHH combined method for improving odontogenic differentiation efficiency of keratinocyte stem cells

Abstract

The invention relates to an FGF8 / SHH double-growth factor combined method for improving differentiation efficiency of keratinocyte stem cells originated from epithelial tissue of an animal into tooth forming enamel-secreting cells. The method comprises the following steps: separating and culturing keratinocyte stem cells so as to obtain cultured keratinocyte stem cells, recombining protein A agarose beads having adsorbed double growth factors, granules of tooth germ epithelium and mesenchymal tissue separated from a molar region of an mouse embryo and the keratinocyte stem cells so as to obtain chimeras, and finally transplanting the chimeras into renal capsule of a nude mouse for induction culture, wherein, the concentration of FGF8 in mixed liquor is 120 to 150 mu g / mL and the concentration of SHH in the mixed liquor is 200 to 250 mu g / mL. The method enables a tooth forming rate and an enamel forming rate to be respectively increased to 48% to 67%; meanwhile, the FGF8 / SHH double-growth factor combined method is simple to operate and facilitates popularization.

Description

technical field [0001] The present invention relates to a method for inducing odontogenic differentiation of epidermal stem cells, in particular to improving the differentiation efficiency of epidermal stem cells derived from animal epithelial tissue to enamel-forming cells by using FGF8 / SHH (fibroblast growth factor 8 / Sonic hedgehog) double growth factor combination method Methods. Background technique Teeth can not only chew food, help pronunciation, but also have a great influence on the face. However, human hereditary tooth defects are a relatively common phenomenon. At the same time, with the growth of age, almost all people have different degrees of tooth defects or even loss. Teeth, especially the enamel part, will not be able to complete repair and regeneration once it is lost. Therefore, the regeneration of human teeth is a research work with great application prospects, social benefits and market value. According to the characteristics of tooth development, t...

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Problems solved by technology

Induction of human keratinocytes into enamel- secreting ameloblasts. Deve-lopmental Biology. 2010, 344:795-799.), but the efficiency of chimeras to form tooth structure (odontogenic rate) is about 28%, in which epidermal stem cells differentiate to form enamel The efficiency (amellation rate) of the cells is about 31%, which is not very efficient

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