(英文)
2. Dissociated spleen cells were suspended with PBS and strained through a cell strainer (BD Biosciences 352340).
3. After centrifuging at 1,000 rpm for 5 min, collected cells were re-suspended in DMEM medium and added to the same volume of lympholyte (Cedarlane), and then centrifuged at 1,000 g for 20 min.
IMPORTANT
(i) The purity of the starting cells is important for achieving STAP conversion. For lymphocytes, contamination with red blood cells may inhibit the reprogramming event. When using adherent cells, the presence of extracellular matrix may interfere with reprogramming.
(ii) Alternatively, red blood cells may be removed by suspension of the cell pellet in 1.8 ml of H2O (Sigma W3500). After 30 seconds, add 0.2 ml of 10× PBS (Gibco 70011-044), followed by 3 ml of 1× PBS (Gibco 10010-023), and strain the cell suspension through a cell strainer.
