Quality and quantity of Asian elephant (Elephas maximus) peripheral blood mononuclear cells isolated by polymer of sucrose or colloidal silica solution gradient centrifugation

Abstract

Peripheral blood mononuclear cells (PBMCs) are useful in cellular immunology, infectious disease diagnosis and molecular genetics research. This study was aimed to determine an appropriated procedure for PBMCs isolation from an Asian elephant (Elephas maximus) by using gradient centrifugations method. The efficacy of four different density solutions including polymer of sucrose solution (Ficoll-Hypaque mixture) of density 1.077 and colloidal silica solution (Percoll) of density 1.077, 1.078 and 1.079 g/cm3 were compared in elephant PBMC separation. Quality and quantity of isolated PBMCs were estimated by using five parameters including cell purity, cell recovery rate, cell viability, viability of PBMCs after 5 days cultivation and total cell count. The results showed that the total cell count was significantly different among 4 treatments, in which Percoll with density 1.079 g/cm3 yielded the highest PBMCs (average = 3.94 X 107 cells/cm3) and highest PBMCs recovery rate (average = 82.61%). However, the purity, viability and viability of PBMCs after 5 days cultivation were not significantly different (P<0.05). In conclusion, Percoll with a density of 1.079 g/cm3 is the most suitable density gradient treatment for Asian elephant PBMCs isolation in both quantity and quality aspects.