Separation of mononuclear bone marrow cells using the Cobe 2997 blood cell separator.
In the present study, we report the results of our evaluation of the use of the continuous-flow cell separator Cobe 2997 to isolate from human bone marrow (BM) aspirates the mononuclear cell (MNC) fraction containing hematopoietic stem cells. This MNC concentrate is isolated in 15% of the original BM volume and contains 23% of the initial nucleated cells. It is enriched as concerns the BM MNC fraction (lymphocytes + monocytes recovery; 80%), whereas the contamination with granulocytes, red blood cells and platelets is reduced to 7.2, 1.5% and 41%, respectively, of the cells initially present in the BM suspensions. Furthermore, it is demonstrated that this MNC concentrate is highly enriched in granulocyte-macrophage-colony-forming cells (CFU-GM; recovery 83%). The method is simple, inexpensive, efficient and reproducible. It allows rapid processing of a large volume of BM without substantial loss of hematopoietic progenitor cells. It represents a valuable method of BM MNC concentration prior to further in vitro manipulations such as T cell or tumor cell depletion or cryopreservation.
Faradji
A
A
CHU Hautepierre, BMT Unit, Strasbourg, France.
Andreu
G
G
Pillier-Loriette
C
C
Bohbot
A
A
Nicod
A
A
Autran
B
B
Bergerat
J P
JP
Rio
B
B
Leblond
V
V
Binet
J L
JL
D000293:Adolescent; D000328:Adult; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation; D002469:Cell Separation / Q000379:methods*; D002648:Child; D002675:Child, Preschool; D006801:Humans