A nylon wool filter coated with human immunoglobulin for rapid depletion of monocytes and myeloid cells from peripheral blood stem cell transplants.
The aim of this study was to develop an inexpensive method for reducing the number of differentiated cells from granulocyte colony-stimulating factor-mobilized leukocytapheresis products (LPs) containing peripheral blood stem cells. Analysis of LPs showed the presence of significant numbers of monocytes and myeloid cells. The myeloid cells represented largely immature stages of the granulocyte lineage (myelocytes and metamyelocytes). We investigated whether these cells could be selectively depleted by filtration over nylon wool. Filtration of LP samples over nylon wool in a medium containing fetal calf serum resulted in variable but on average low yields of CD34+ cells (48 +/- 30%; n=13) and strongly variable depletions of myeloid cells. The adherence of CD34+ cells to the polyamide fiber was partially mediated by activated platelets that were present in the LPs. Removal of platelets by counterflow centrifugal elutriation before filtration resulted in increased yields of CD34+ cells in the filtrates (65 +/- 13%; n=10). The yield of progenitor cells was similarly enhanced when trisodium citrate, a chelating substance, was added to the filtration medium. Adherence of the myeloid cells to the nylon fiber was promoted by preincubation of the columns with human immunoglobulin (Ig) (2 mg/mL). Small-scale filtrations of LP samples in the presence of trisodium citrate over columns with Ig-coated nylon wool resulted in removal of 96 +/- 4% of the monocytes and 74 +/- 18% of the myeloid cells, with a yield of 71 +/- 15% CD34+ cells and 67 +/- 10% granulocyte-monocyte colony-forming units (CFU-GM) (n=23). There was no loss of primitive stem cells during the procedure: the yield of late-appearing cobblestone area-forming cells (CAFCs, week 6) was 110 +/- 30% (n=4). CFU-GM production per CAFC-derived clone was unchanged upon filtration, indicating that the quality of stem cells was not affected. Moreover, the proportions of CD34+ cells expressing a primitive immunophenotype (CD38low or Thy-1+) were unchanged after filtration. Further enrichment of progenitor cells was obtained by separation of LP samples by elutriation before filtration. The combination of these two techniques resulted in complete removal of platelets, 89 +/- 7% depletion of erythrocytes, and 91 +/- 6% reduction of leukocytes, with a 50% yield of CD34+ cells (n=14). In conclusion, we have developed a rapid filtration technique by which monocytes and myeloid cells can be depleted from LP samples, with only minor loss of colony-forming cells and complete recovery of primitive stem cells.
Kwekkeboom
J
J
Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, University of Amsterdam.
Buurman
D E
DE
Ploemacher
R E
RE
Baars
J W
JW
Loos
H A
HA
Slaper-Cortenbach
I C
IC
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis / Q000276:immunology; D001781:Blood Component Removal / Q000379:methods*; D001792:Blood Platelets / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology*; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D019343:Citric Acid; D004358:Drug Therapy; D005312:Fetal Blood; D005374:Filtration / Q000379:methods; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation / Q000295:instrumentation*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D007136:Immunoglobulins / Q000494:pharmacology; D007937:Leukapheresis; D009000:Monocytes / Q000166:cytology*; D009757:Nylons; D013997:Time Factors
D018952:Antigens, CD34; D007074:Immunoglobulin G; D007136:Immunoglobulins; D009757:Nylons; D016179:Granulocyte Colony-Stimulating Factor; D019343:Citric Acid