LPS priming of stromal cells to generate LPS-specific exosome educated macrophages
The disclosure relates to an ex vivo generated population of educated macrophages specific to LPS and methods of making and using such macrophages.
1. A method for generating an educated macrophage, the method comprising the steps of:
isolating extracellular vesicles from a mesenchymal stem cell previously exposed to lipopolysaccharide (LPS), and
co-culturing a CD14+ cell with the extracellular vesicles in vitro until the CD14+ cell acquires an anti-inflammatory macrophage phenotype, wherein the mesenchymal stem cell is exposed to about 800 ng/ml to about 1200 ng/ml LPS.
2. The method of claim 1 , wherein the CD14+ cell and the extracellular vesicles are co-cultured for at least 2 days.
3. The method of claim 1 , wherein the mesenchymal stem cell is exposed to LPS for at least 2 hours.
4. The method of claim 1 , wherein the CD14+ cell is a macrophage.
5. The method of claim 1 , wherein the CD14+ cell is a monocyte and wherein the CD14+ monocyte and the extracellular vesicle are co-cultured for at least 5 days.
6. A population of anti-inflammatory macrophages produced by the method of claim 1 , wherein the anti-inflammatory macrophage phenotype is characterized as FLT-3L high, IL-15 high, CD73 high, CD86 low, and HLA-DR low as compared to control macrophages.
7. A method for generating an educated monocyte, the method comprising the steps of:
isolating extracellular vesicles from a mesenchymal stem cell previously exposed to lipopolysaccharide (LPS), and
co-culturing a CD14+ monocyte with the extracellular vesicles in vitro until the CD14+ monocyte acquires an anti-inflammatory monocyte phenotype, wherein the mesenchymal stem cell is exposed to about 800 ng/ml to about 1200 ng/ml LPS.
8. The method of claim 7 , wherein the CD14+ monocyte and the extracellular vesicles are co-cultured for at least 2 hours.
9. The method of claim 8 , wherein the CD14+ monocyte and the extracellular vesicles are co-cultured for at least 24 hours.
10. The method of claim 7 , wherein the mesenchymal stem cell is exposed to LPS for at least 12 hours.
11. A population of anti-inflammatory monocytes produced by the method of claim 7 , wherein the anti-inflammatory monocyte phenotype is characterized as PD-L1 high, CD206 low, CD163 low, IL-15 high, CD73 high, CD86 low, CD16 low and IL-6 high as compared to control monocytes.