Viability assessment of fresh chicken blastodermal cells by flow cytometry

Abstract

This study describes the viability assessment of fresh chicken blastodermal cells (BCs) by flow cytometry using a novel DNAdetecting far-red-fluorescing dye, DRAQ5 and LIVE/DEAD (LD) Fixable Green Dead Cell Stain Kit. Overall, the technique provides both quantitative and qualitative evaluation of BCs. Particular attention was put on the determination of a portion of nucleated (DRAQ5+ ) and dead (DRAQ5+ /LD+ ) BCs from freshly isolated samples. The results showed that the proportion of nucleated cells from a sample was ranged from 50.39 to 72.58 % and dead cells from 0.40 to 11.38 % in all repetitions. Furthermore, our study suggests that flow cytometry allows precise examination of fresh BCs viability in a very short time and that a combination of DRAQ5/LD staining is suitable for the detection of nuclear BCs and their viability evaluation, respectively. Moreover, this technique might also be applied for the evaluation of frozen-thawed BCs after cryopreservation process for animal gene bank.