Abstract:

BACKGROUND: The culture of human periodontal ligament cell (hPDLC) is the basis of biological research of periodontal cells. The success rate of primary culture of periodontal membrane cells is low, which restricts the biological research of periodontal cells. How to seek a simple effective culture method or how to elevate success rate of primary culture is always one of hot focus studies of biology of periodontal cells.
OBJECTIVE: To explore an effective culture technique of hPDLC by comparing the effects of tissue explant, enzymatic digestion, the combination of tissue explant and enzymatic digestion.
METHODS: The tissues of periodontal ligament isolated from 58 normal human permanent teeth were collected and divided into three groups randomly, and then primarily cultured using tissue explant, enzymatic digestion, the combination of tissue explant and enzymatic digestion, followed by subculture. Cell growth and morphology were observed under an inverted microscope. Vimentin and keratin were identified by immunohistochemistry. The fourth passage of cells were collected to draw growth curve.
RESULTS AND CONCLUSION: The successful rate of the combination of tissue explant and enzymatic digestion (35%) was significantly higher than others (11% and 21%). The morphological and biological characteristics of the hPDLC in present study were similar to those typical hPDLC, which grew well. The growth curve of the fourth passage of cells was similar to “S” shape, with significant detention period, logarithmic phase and plateau phase. Results have suggested that the combination of tissue explant and enzymatic digestion is relatively simple and has a high successful rate, so it can be regarded as an effective method for hPDLC culture.