Isolation of murine hepatic lymphocytes using mechanical dissection for phenotypic and functional analysis of NK1.1+ cells

Figure 1 Effects of enzymatic digestion on surface molecules of hepatic lymphocytes. ^aP < 0.05, NK1.1 vs other groups except DX5; ^bP < 0.01, DX5 vs other groups except NK1.1.

Figure 2 Cell yields between the two isolating methods. Hepatic lymphocytes were isolated using mechanical dissection method and the enzymatic digestion method, respectively, from normal C57BL/6 mice (A), Poly (I:C)-treated C57BL/6 mice (B) or ConA-treated C57BL/6 mice(C).

Figure 3 Hepatic lymphocytes isolated with mechanical dissection were suitable for phenotypic analysis of NK1. 1⁺ cells. A: Lymphocytes from liver, lung, spleen and thymus of normal C57BL/6 mice were labeled with two-color immunofluorescence; B, C: C57BL/6 mice were injected with PBS, Poly (I:C) or ConA, respectively. Hepatic lymphocytes were isolated and labeled for phenotype (CD3 and NK1.1) and intracellular cytokine (IFN-γ and TNF-α) detection.

Figure 4 Hepatic lymphocytes isolated with mechanical dissection were suitable for the functional analysis of NK1. 1⁺ cells. A: Viability of hepatic lymphocytes isolated with two methods; B: Cytotoxicity analysis of hepatic lymphocytes. Hepatic lymphocytes were isolated by two different methods, respectively, from control B6 mice (a) or Poly (I:C)-treated B6 mice (b). Their cytotoxicity against YAC-1 cells was tested at the (E/T) ratios.