CFU-c enrichment from human bone marrow using a discontinuous Percoll gradient and soybean agglutinin in comparison with Ficoll-paque

Abstract

This study compares the efficiency of different methods of separation of human bone marrow in vitro prior to transplantation. Separation on Ficoll-paque resulted in a median recovery of 11.3 X 10(4) CFU-c/10(9) nucleated cells harvested. The recovery from the major CFU-c containing fraction following separation on a discontinuous Percoll gradient was 10.8 X 10(4) CFU-c/10(9) nucleated cells. The CFU-c recovered from Percoll were contained in a smaller number of cells, resulting in enrichment for CFU-c of 1.6 times that of Ficoll-paque. Further CFU-c enrichment to approximately three times that of Ficoll-paque separation was possible by treatment of Percoll fractionated cells with soybean agglutinin (SBA). However this caused an overall loss of 67% of CFU-c. Cells remaining after SBA treatment of the CFU-c containing Percoll fraction had increased spontaneous DNA synthesis and decreased PHA responses. There was no significant change in the mixed lymphocyte reaction in comparison with Ficoll-paque.