Fetal bovine serum-derived exosomes regulate the adipogenic differentiation of human bone marrow mesenchymal stromal cells in a cross-species manner
Fetal bovine serum (FBS) containing a large amount of exosomes that can interfere with the analysis of exosomes derived from cultured cells. We investigated the effect of exosomes derived FBS (FBS-Exos) on adipogenic differentiation of human bone marrow mesenchymal stromal cells (HBM-MSCs) and the underlying molecular mechanisms.
FBS-Exos absorption by HBM-MSCs can be detected by laser scanning confocal microscope, and the treatment of exosomes resulted in a decrease in lipid droplet formation and reduced expression of genes associated with adipogenic differentiation of HBM-MSCs. miR-1246 was identified as microRNA is abundant in FBS-Exos by public identification data sequencing and RT-qPCR validation. In addition, miR-1246 in the HBM-MSCs excess causes decreased levels of adipogenic differentiation, whereas miR-1246 in FBS-Exos knockdown attenuated inhibitory effects on adipogenic differentiation.
Our results indicate that FBS-Exos inhibits adipogenic differentiation of HBM-MSCs by means of cross-species and miR-1246 transferred by FBS-Exos partly contribute to this effect.
Fetal bovine serum-derived exosomes regulate the adipogenic differentiation of human bone marrow mesenchymal stromal cells in a cross-species manner
Human Cells Grow With or Without Replacement for Fetal Bovine Serum
security problems on cell-derived pharmaceutical products manufactured in the supplement of fetal bovine serum (FBS) has fueled the demand to replace FBS.
Here, four new alternative to FBS marketed compared: xeno-free product called Ess® cells, human platelet lysates marketed as GroPro®, and two mixtures of serum adult cattle vary in their proportion of neonatal growth factor, called Liporo® and FetalGro®. Endothelial cell line (C2BBe1) and neural cell lines (SHSY5Y) near confluency media with 10% FBS is selectively etched and taken through a step-wise algorithm 25 days to replace FBS, and other human endothelial cell line (HRA- 19) studied for C2BBe1 emulate. Cells were stained and counted and compared to the survival, migration, and spheroids.
The C2BBe1 and HRA-19 cell line failed to proliferate in 10% of cells Ess® but growing at 10% or 10% FetalGro® GroPro® quite good compared with 10% FBS reference. The SH-SY5Y, just FetalGro® FBS efficacy of this approach. These are all inferior to 11 different branded many of FBS (positive control), but five days to switch only between brands FBS, 4 out of 11 supported less proliferation of endothelial FBS reference in HRA-19 (p <0.004). In addition, the neurospheres were enriched in two many brands of FBS and FetalGro® (each p <0.004), neurospheres be undesirable phenotype for nerve cell applications. Because GroPro® platelet-derived growth supplement prominent among non-FBS to allow proliferation without going spheroids, it seems the best (be aware that the cells still grew more slowly in it compared to FBS).
Description: Protein A is a 42 kDa surface protein originally found in the cell wall of the bacterium Staphylococcus aureus. It can bind immunoglobulins.
Description: Protein A is a 42 kDa surface protein originally found in the cell wall of the bacterium Staphylococcus aureus. It can bind immunoglobulins.
Description: Protein A is a 42 kDa surface protein originally found in the cell wall of the bacterium Staphylococcus aureus. It can bind immunoglobulins.
Description: Protein A is a 42 kDa surface protein originally found in the cell wall of the bacterium Staphylococcus aureus. It can bind immunoglobulins.
Although there is no perfect substitute is found among alternative to FBS, the algorithm for the switch to be useful in future testing new alternative to FBS as a need to switch from FBS and expanding pharmaceutical products safety for human use. Keywords: C2BBe1; SH-SY5Y; fetal bovine serum; neurospheres; xeno-free.
