Figure 8.

Design of experiment to determine activity of different part of HDSN. (A) Scheme used to collect HDSN and exosome. After removal of cells and cell debris the supernates were ultracentrifuged and the upper (representing supernate devoid of exosome), middle part (representing supernate devoid of exosome), and pellet (representing small extracellular vesicle = exosome) were collected. The pellet was further processed as shown in (B) to obtain more pure exosome. All the fractions were used to determine if they could stimulate the LD ALL3 cells or not. (B) Steps for the exosome purification procedure based on differential ultracentrifugation and sucrose gradients. The right side of the arrow indicates speed, time or washing solution used for each step. Pellets (cells, dead cells, cell debris) were discarded after each of the first three steps, and the supernatant was ultracentrifuged. Then the pellet was washed, and also purified using sucrose gradient and washed again with PBS. The washes were discarded and the pellet was incubated at -20°C until further use.