Aim: To optimize the traditional magnetic activated cell sorting (MACS) so as to enhance the purification and keep the viability of cells after separation.
Methods: Take stem cell antigen-1 labeled hemopoietic stem/progenitor cells (Sca-1(+);HSC/HPC) for example. The traditional MACS and the optimized MACS were applied to obtain the Sca-1(+); HSC/HPC from mouse bone marrow respectively. The purifications of Sca-1(+); cells from two groups were tested by flow cytometry; The survival rates of Sca-1(+); cells from two groups were detected by trypan blue dye; The proliferation of Sca-1(+); cells was detected by cell counting kit-8(CCK-8); the differentiation capacity of Sca-1(+); cells was measured by CFU-Mix.
Results: The purification of Sca-1(+); cells was up to 93% by the optimized MACS compared to 87% by the traditional one; the recovery rate of positive cells was 73% by the optimized MACS compared to 62.3% by the traditional one; there was no statistical difference in the viability and proliferation of Sca-1(+); cells and the capacity of producing CFU-Mix between two groups.
Conclusion: The optimized MACS can significantly enhance the purification and recovery rate of Sca-1(+); cells without influence of cell viability and biological features, which can be considered to apply in other MACS.