Active Leukemia Inhibitory Factor (LIF)

Leukemia inhibitory factor (LIF), is an interleukin 6 class cytokine that affects cell growth by inhibiting differentiation. LIF as a cytokine also has another fuction including: the growth promotion and cell differentiation of different types of target cells, influence on bone metabolism, cachexia, neural development, embryogenesis and inflammation. The activity of LIF is usually measured by a cell proliferation assay using TF-1 cells. TF-1 cells were seeded into triplicate wells of 96-well plates at a density of 20000 cells/well with 10% serum standard 1640 which contains various concentrations of recombinant human LIF. After incubated for 3 days, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10 µl of CCK-8 solution was added to each well of the plate, then the absorbance at 450 nm was measured using a microplate reader after incubating the plate for 2-4 hours at 37 ℃. Proliferation of TF-1 cells after incubation with LIF for 3 days observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with rhLIF for 3 days. The result was shown in Figure 2. It was obvious that rhLIF significantly increased cell viability of TF-1 cells. The ED50 is 1.86 ng/ml.