The invention relates to the field of
medical biology, and provides a separation and amplification method and application of human
dental pulp stem cells. The separation and amplification method comprises the following steps: S1, taking human dental pulp tissue,
cutting the human dental pulp tissue into pieces, and adding
collagenase I,
hyaluronidase and
dispase II for
digestion to obtain digestedtissue and
cell suspension; S2, filtering the digested tissue and
cell suspension by using a filter screen, centrifuging the filtrate, and taking
cell precipitate to obtain human
dental pulp stem cells; and S3, inoculating the human
dental pulp stem cells into a culture dish pre-coated with human
fibronectin, adding a culture solution for culture, and performing subculture when convergence rate reaches 70-80%. The method has a short time for cell adherence, number of cells cultured to the 10th day of at least 4.5x105,
high cell purity, positive expression rate of dental pulp
stem cell surfacemarkers CD73, CD90 and CD105 above 99%, and negative index expression rate lower than 1.2%.