Isolation and biological characteristics of exosomes derived from periodontal ligament stem cells

doi:10.19439/j.sjos.2019.04.002

Abstract

Abstract: PURPOSE: To isolate and identify exosomes derived from periodontal ligament stem cells (PDLSCs) collected by ultracentrifugation. METHODS: Using the limiting dilution technique, human PDLSCs were isolated and expanded. The cell culture supernatant of PDLSCs was collected. Exosomes were collected and purified with a ultracentrifugation method. Biological characteristics of exosomes derived from PDLSCs were measured by transmission electron microscopy (TEM), Western blot and nanosight tracing analysis (NTA). RESULTS: Exosomes could be successfully isolated from the supernatant of PDLSCs by a ultracentrifugation method. Under TEM, the PDLSC-derived exosomes exhibited elliptic or saucer-like shape and the central area had lower electron density than the circum area. The PDLSC-derived exosomes could express the common surface adhesion molecules CD9, CD63, CD81 and TSG101. NTA results showed that the collected exosomes had a size around (119±12.1) nm and an approximate concentration of (3.80±0.39)×10⁸ particles/mL. CONCLUSIONS: Exosomes derived from PDLSCs can be collected by a ultracentrifugation method, which expresses common membrane proteins and morphological characteristics of exosomes.

Key words: PDLSCs, Exosomes, Extracelluar vesicles, Ultracentrifugation

CLC Number:

R783.1

ZHAO Li-ru, MAO Jia-qi, ZHAO Bing-jiao, CHEN Jing. Isolation and biological characteristics of exosomes derived from periodontal ligament stem cells[J]. Shanghai Journal of Stomatology, 2019, 28(4): 343-348.

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