title: Preparation of autologous bone marrow grafts for cryopreservation using the AS104 cell separator.
abstract: We evaluated the AS104 cell separator (Fresenius AG, Bad Homburg, Germany) for ex vivo processing of bone marrow (BM) grafts of 43 patients suffering from germ cell cancer (GCC, n = 22), acute lymphocytic leukemia (ALL, n = 13) and malignant lymphoma (ML, n = 8). Recoveries of total nucleated cells (TNC), mononuclear cells (MNC) and colony-forming units granulocyte-macrophage (CFU-GM) were determined in the BM concentrates prepared for cryopreservation. Hematopoietic reconstitution was analyzed in patients who underwent autologous transplantation following high-dose radio-/chemotherapy (HDRCT). Processing of the BM suspension with a median volume of 1,013 ml (range: 422-1,574) resulted in 156 ml (80-186) within 50-120 min (median: 90). In the BM concentrates, medians of 28.6% TNC (10.6-69.6), 37.9% MNC (22.3-86.4), and 52.4% CFU-GM (20.8-96.4) were recovered. Twenty-six patients underwent HDRCT with reinfusion of autologous BM and were evaluable for engraftment. They received a median of 0.8 x 10(8) MNC/kg (0.3-1.6 x 10(8)) and 2.2 x 10(4) CFU-GM/kg (0.6-12.8 x 10(4) for hematopoietic rescue. Engraftment with neutrophils > 500/microliter occurred in a median time of 12 days (8-33) in all patients. We conclude that ex vivo processing of autologous BM with median recovery rates of 37.9% for MNC, and 52.4% for CFU-GM, results in a cell population that can rescue patients from HDRCT. The described technique is convenient, time-efficient, and provides reliable results in preparing BM autografts for cryopreservation.
authors.affiliation: Department of Internal Medicine, Virchow Clinic, Humboldt University, Berlin, Germany.
authors.forename: O
authors.initials: O
authors.lastname: Rick
authors.forename: H G
authors.initials: HG
authors.lastname: Heuft
authors.forename: G
authors.initials: G
authors.lastname: Wittmann
authors.forename: R
authors.initials: R
authors.lastname: Blasczyk
authors.forename: D
authors.initials: D
authors.lastname: Huhn
authors.forename: N
authors.initials: N
authors.lastname: Schwella
mesh_terms: D000293:Adolescent; D000328:Adult; D016026:Bone Marrow Transplantation; D002469:Cell Separation / Q000295:instrumentation; D002648:Child; D002675:Child, Preschool; D015925:Cryopreservation; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D008875:Middle Aged