GSE113634
Transcriptome profiling of osteoblasts treated with uMSC-derived exosomes
To investigate the function and affected pathways of uMSC-derived exosomes on ossification, we used human and rat osteoblast cell line hFOB1.19 and ROS1728 treated with either exosomes or exosome-free supernatent (which is the  supernatent of exosome pelleting after ultracentrifugation) and a non-treated group as negative control. We take advantages of high through-put sequencing technology to fully reveal the transcriptome changes of the treated cells. Overall design: we used human and rat osteoblast cell line hFOB1.19 and ROS1728 for the study. For exosome collection, umbilical cord were collected and washed with PBS and subjected to primary culture immediately. The samples were first depleted of blood vessels and minced to small pieces then put to culture flasks for primary culture. 10%FBS high-glucose DMEM were used as culture medium for primary cell culture. When cells reach 80% confluent, the cell were lysed with trypsin for passage. After 2 passages, the primary cells were ready to perform exosome collection. For exosome collection, we use 1% exosome-free FBS/DMEM for culture, the medium were collected at a 3-days interval, and the collected culture medium of  were subsequently subjected to ultracentrifugation. After the ultarcentrifugation, the pellet were washed with PBS and ultarcentrifugated again before use. The exosomes were measured for protein concentration using standard method, and 100ng/ml exosomes were used to treat the samples. 48h after treatment, the cells were collected for total RNA extraction.