Multi-color flow cytometry bovine AT-MSCs

In this study, we aimed to immunophenotype bovine adipose tissue (AT)-derived MSCs using multi-color flow cytometry. 13 commercial Abs were screened for recognizing bovine epitopes using the appropriate positive controls.
Conclusion:
Using flow cytometry and immunofluorescence microscopy, cross-reactivity was confirmed for CD34, CD73, CD79α and CD90. Unfortunately, none of the evaluated CD105 and CD106 Abs cross-reacted with bovine cells. Subsequently, AT-derived bovine MSCs were characterized using multi-color flow cytometry based on their expression of nine markers. Bovine MSCs clearly expressed CD29 and CD44, and lacked expression of CD14, CD45, CD73, CD79α and MHCII, while a variable expression was observed for CD34 and CD90. In addition, the mRNA transcription level of different markers was analyzed using RT-qPCR. For daily validation of the flow cytometer, CytoFlex Daily QC Fluorospheres (Beckman Coulter, B53230) are used. For weekly validation of the flow cytometer, an 8-peak validation is performed using rainbow calibration beads (Spherotech, RCP-30-5A).