Influence of albumin and placement time outside the incubator on viability of isolated cytokine-induced killer cells: a simple method for detection / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To study the influence of albumin supplementation in the medium and the placement time outside the incubator on the viability of isolated cytokine-induced killer (CIK) cells.</p><p><b>METHODS</b>CIK cells labeled with anti-CD3-FITC and anti-CD56-PE or with FQ-AE were observed under fluorescence microscope. The effect of albumin in the cell medium on the cell viability was analyzed using flow cytometry with Annexin V-FITC after different time lengths of placement.</p><p><b>RESULTS</b>The clones of CIK cells from the patient with esophageal carcinoma were small and scattered in the medium, but the clones from the patients with pancreatic cancer were large and densely distributed. CD3(+)CD56(+) cells could be detected under fluorescence microscope. The addition of albumin in the medium did not obviously affect cell apoptosis and death of CIK cells. CIK cells placed outside the incubator for less than 90 min showed a significant lower apoptosis rate than the cells placed for 150 min, whereas the cell death rate did not vary significantly with the placement time.</p><p><b>CONCLUSION</b>CIK cells from different cancer patients present with different growth pattern of the cells clones. Labeling with anti-CD3-FITC and anti-CD56-PE allows convenient counting of the newly generated CD3(+)CD56(+) CIK cells and FQ-AE labeling can be used for quantitative assessment of cell death. Albumin is not necessary in the medium of CIK cells. Prolonged placement (for over 90 min) of CIK cells outside the incubator should be avoided, and the placement time should be shorten as much as possible.</p>