title: Analysis for recovery and loss of mononuclear cells and colony-forming units granulocyte-macrophage during ex vivo processing of autologous bone marrow.
abstract: During ex vivo processing of autologous bone marrow (BM) substantial loss of stem and progenitor cells should be avoided to achieve rapid and sustained hematopoietic reconstitution after high-dose radio-/chemotherapy. We processed 25 autologous BM grafts with the Fresenius AS104 cell separator for cryopreservation and we determined recoveries for mononuclear cells (MNC) and colonyforming units granulocyte-macrophage (CFU-GM) in the BM concentrates. To identify cell loss in BM fractions not cryopreserved, we investigated the MNC and CFU-GM content of BM fat and BM blood. MNC and CFU-GM recovery yielded a mean ( +/- SEM) of 42 +/- 12 and 54 +/- 20% in the BM concentrate. BM fat showed a mean loss of 7 +/- 5% for MNC and 4 +/- 3% for CFU-GM, BM blood 30 +/- 12% for MNC and 13 +/- 13% for CFU-GM, respectively. CFU-GM recovery was significantly higher in the BM concentrate of patients with hematologic malignancy (HM) compared with patients suffering from germ cell cancer (GCC): 66 +/- 21 vs. 43 +/- 12% (p < 0.02). Seventeen patients (7 GCC, 10 HM) underwent high-dose chemotherapy or radio-/chemotherapy and were autografted with 0.8 +/- 0.2 x 10(8) MNC/kg and 3.7 +/- 2.0 x 10(4) CFU-GM/kg. All patients achieved engraftment with neutrophils > 0.5 x 10(9)/l at a mean of 14 +/- 6 days. We conclude that: (1) ex vivo processing of autologous BM with a mean of recovery of 42% for MNC and 54% for CFU-GM in the BM concentrate can result in a cell population capable of sustained hematopoietic reconstitution, (2) CFU-GM recovery is significantly higher in patients with HM than in patients with GCC and (3) 37% MNC and 17% CFU-GM represent in fact cell losses recovered from BM fractions not cryopreserved (BM fat, BM blood). Furthermore, it is likely that MNC and CFU-GM not recovered from BM concentrate, BM fat and BM blood are cell losses related to the cell separator.
authors: Virchow-Klinikum, Humboldt-Universität Berlin, Deutschland.
authors: N
authors: N
authors: Schwella
authors: H G
authors: HG
authors: Heuft
authors: O
authors: O
authors: Rick
authors: R
authors: R
authors: Blasczyk
authors: G
authors: G
authors: Wittmann
authors: D
authors: D
authors: Huhn
mesh_terms: D000293:Adolescent; D000328:Adult; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation / Q000379:methods*; D002648:Child; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000627:therapeutic use; D006098:Granulocytes / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006689:Hodgkin Disease / Q000628:therapy; D006801:Humans; D007700:Kinetics; D008228:Lymphoma, Non-Hodgkin / Q000628:therapy; D008264:Macrophages / Q000166:cytology*; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology*; D009373:Neoplasms, Germ Cell and Embryonal / Q000628:therapy; D017063:Outcome Assessment, Health Care; D010976:Platelet Count; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000628:therapy; D014182:Transplantation, Autologous
chemical_list: D016179:Granulocyte Colony-Stimulating Factor