Separation of rat bone marrow cells by counterflow centrifugal elutriation: a model for studying the effects of lymphocyte depletion.
We have developed a simple three flow-rate, fixed rotor speed, counterflow centrifugal elutriation (CCE) procedure that permits the isolation of an engraftable lymphocyte-depleted (greater than 98%) fraction from ACI rat bone marrow. The different cell fractions were characterized by morphology, alloreactivity in mixed lymphocyte culture and limiting dilution analysis, colony-forming capacity, and their capacity to reconstitute hematopoiesis and effect a graft-versus-host reaction in lethally irradiated allogeneic hosts. After CCE fractionation of ACI rat marrow, transplantation of the lymphocyte-depleted marrow fraction resulted in sustained engraftment without evidence of clinical or histologic acute graft-versus-host disease (GVHD). CCE fractionation of rat bone marrow may be a useful preclinical model for studying lympho-hematopoietic and immune reconstitution after transplantation with lymphocyte-depleted donor marrow, as well as for studying the role of lymphocyte subpopulations on engraftment, acute GVHD, and leukemia relapse in syngeneic and allogeneic bone marrow transplantation.
Wagner
J E
JE
Department of Oncology, Johns Hopkins Hospital, Baltimore, MD 21205.
Donnenberg
A D
AD
Noga
S J
SJ
Wiley
J M
JM
Yeager
A M
AM
Vogelsang
G B
GB
Hess
A D
AD
Santos
G W
GW
D000818:Animals; D001854:Bone Marrow Cells; D002469:Cell Separation; D002498:Centrifugation / Q000379:methods; D004195:Disease Models, Animal; D006086:Graft vs Host Disease / Q000517:prevention & control; D008212:Lymphocyte Depletion; D051381:Rats; D011917:Rats, Inbred Lew; D011919:Rats, Inbred Strains