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Figure 1. ���շ��ѱ�����™ Mouse Spleen Dissociation Kit Achieves High Cell Viability and Yield

Mouse spleen tissue was processed into single-cell suspension using the ���շ��ѱ�����™ Mouse Spleen Dissociation Kit and the ���շ��ѱ�����™ Tissue Dissociator, an alternative automated system, or a manual dissociation method. (A) Viability of total nucleated cells. (B) Yield of viable cells per whole spleen tissue (63 - 152 mg). (C) Yield of spleen myeloid and lymphoid subsets. Viability, yield, and subset composition were assessed by flow cytometry. Red blood cells were lysed with ammonium chloride solution before analysis. Data are presented as mean ± SD (n = 17 - 26). * p < 0.05, one-way ANOVA with Tukey's multiple comparisons test.

Figure 2. ���շ��ѱ�����™-Processed Mouse Splenic T Cells Proliferate Upon Activation

Mouse spleen tissue was processed into single-cell suspensions using the ���շ��ѱ�����™ Mouse Spleen Dissociation Kit and the ���շ��ѱ�����™ Tissue Dissociator. (A) Splenic T cells were then isolated using EasySep™ Mouse T Cell Isolation Kit. T cells were seeded at 2 x 105 cells in 200 µL of ImmunoCult™-XF T cell Expansion Medium and activated with ImmunoCult™ Mouse T Cell Activators in the presence of IL-2 (50 U/mL) for 4 days. (B) Fold expansion of T cells cultured in medium alone or in the presence of ImmunoCult™ activators CD3/CD28 or CD3/CD28/CD2. Flow cytometry analysis of T cells showing (C) the expression of T cell activation marker CD25 on Day 0 and Day 1 after activation and (D) the proliferation of CellTrace™ Violet-labeled T cells on Day 0 and Day 4 of activation. (E) Representative light microscopy images of T cell cultures on Days 0 and 4 after activation. Data are presented as mean ± SD (n = 4).

Figure 3. ���շ��ѱ�����™-Processed Mouse Splenic Monocytes Are Phagocytic and Produce Cytokines upon Activation

Mouse spleen tissue was processed into single-cell suspensions using the ���շ��ѱ�����™ Mouse Spleen Dissociation Kit and the ���շ��ѱ�����™ Tissue Dissociator, an alternative automated system, or a manual dissociation method. (A) Lineage- (CD3, CD45R, CD117, CD49b, Siglec F) CD11b+ monocytes were isolated from the single-cell suspensions using EasySep™ Mouse Monocyte Isolation Kit. (B) The cells were incubated for 2 hours in the presence of pHrodo™ Green-conjugated E. coli BioParticles™ at 2 - 8°C (Cold) or 37°C. The fluorescence of phagocytosed BioParticles™ was measured by flow cytometry. (C) Intracellular flow cytometry staining of TNF-ɑ production by monocytes cultured overnight in the presence of 3 µg/mL Brefeldin A and treated with (+) or without (-) 100 ng/mL of lipopolysaccharides (LPS). Data are presented as mean ± SD (n = 3 - 4).