On the other hand, for the islets co-encapsulated with PTX and then exposed to lymphocytes solution till 7 days, the external environment was totally transparent, indicating that PTX can easily suppress lymphocytes attack on encapsulated islets.
Centrifugation with Ficoll-Hypaque lymphocyte isolation solution (TBD Science, China) was used to isolate splenocytes of immunized mice, according to the manufacturer's instructions.
Lymphocytes from the spleens were isolated by centrifugation in gradient lymphocyte isolation solution for mice (Tianjin Hao Yang Biological Manufacture Co., Ltd., China) at room temperature and washed twice with PBS.
Buffy coats were diluted 1 2 in PBS, and cells were separated in 5 mL Ficoll gradient (lymphocyte isolation solution, Rafer) by centrifugation at 2000 × g for 30 min. PBMCs were collected and washed twice with cold PBS.
PBMC were obtained by density gradient centrifugation using a lymphocyte separation solution.
Mononuclear cells were isolated by density gradient centrifuge using lymphocyte separation solution (Ficoll1.077 g/mL from TBD Biotech, Tianjing, China).
PBMCs were isolated from heparinized blood diluted at a 1 2 ratio with supplemented medium and separated via centrifugation using a lymphocyte separation solution (Lonza).
Peripheral blood mononuclear cells were collected from heparinized blood samples through density gradient centrifugation with lymphocyte separation solution (Nacalai Tesque, Kyoto, Japan).
PBMCs were isolated from local and systemic blood samples obtained from all subjects by Ficoll-Paque density gradient centrifugation and lymphocyte separation solution (Nacalai Tesque).
Monocyte-derived macrophages (MDMs) were obtained from peripheral blood mononuclear cells (PBMCs) that were prepared by centrifugation of whole heparinized venous blood diluted 1/1 with RPMI 1640 (Lonza, Walkersville, MD, USA) over a Lymphocyte separation solution (Lonza) gradient.
