Separation of G-CSF-mobilized PBSC transplants by counterflow centrifugal elutriation: modest enrichment of CD34+ cells but no loss of primitive haemopoietic progenitors.
The suitability of counterflow centrifugal elutriation (CCE) for reduction of the number of non-stem cells in autologous G-CSF-mobilized peripheral blood stem cell (PBSC) transplants was investigated. By cell size-monitored CCE, small cells could be rapidly separated from the haemopoietic progenitor cells present in leukapheresis product (LP) samples. The large cell fraction contained an average 86 +/- 25% of the CD34+ cells and 76 +/- 20% of the granulocyte-macrophage progenitors (CFU-GM) loaded into the separation chamber, and was depleted of 75 +/- 18% of the lymphocytes, 89 +/- 7% of the erythrocytes and 98 +/- 2% of the platelets (n = 21). Due to the presence of high numbers of large immature myeloid cells, which co-elutriated with progenitor cells, enrichment of CD34+ cells in the large cell fraction was only modest (average 1.8 times). No indication of preferential co-elutriation of primitive stem cells with the small cells was obtained. There was no difference in expression of CD38 or Thy-1 on CD34+ cells between the two elutriation fractions. Frequencies of cobblestone-area-forming cells (CAFC) week 6, which are considered to represent cells with long-term repopulating ability, were reduced in the small cell fractions as compared to those in the unseparated samples and the large cell fractions. On average, 100% of CAFC week 6 were recovered in the large cell fractions (n = 5). In conclusion erythrocytes, platelets and 40-50% of leucocytes can be depleted from G-CSF-mobilized PBSC samples by CCE with an almost complete recovery of both clonogenic and primitive stem cells.
Kwekkeboom
J
J
Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, University of Amsterdam.
Buurman
D E
DE
van Hennik
P B
PB
Ploemacher
R E
RE
Loos
H A
HA
Slaper-Cortenbach
I C
IC
D018952:Antigens, CD34; D002469:Cell Separation / Q000379:methods*; D048429:Cell Size; D002498:Centrifugation; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells; D006801:Humans; D016130:Immunophenotyping; D007937:Leukapheresis; D014407:Tumor Cells, Cultured
D018952:Antigens, CD34; D016179:Granulocyte Colony-Stimulating Factor