title: Flow cytometry using annexin V can detect early apoptosis in peripheral blood stem cell harvests from patients with leukaemia and lymphoma.
abstract: Quantifying progenitor cells in peripheral blood stem cell (PBSC) harvests by flow cytometric enumeration of CD34+ cells does not account for cell viability. Cell membrane asymmetry in early apoptosis exposes phosphatidylserine on the cell surface. This can be detected by staining with annexin V FITC. Apoptosis in 30 autologous PBSC harvests mobilised by cyclophosphamide + G-CSF or standard chemotherapy + G-CSF was analysed immediately after collection by dual-colour flow cytometry with CD34 PE and annexin V FITC. Harvests contained a median of 3.4 x 10(6)/kg (range 0.3-91.8) CD34+ cells. Of these 87.6% (range 30-96.5) were annexin V-. In 10% of harvests more than 50% of CD34+ cells were apoptotic. Differences in PBSC mobilisation or collection could not explain the variation in annexin V binding. Cyclophosphamide + G-CSF significantly increased the yield of CD34+ cells but did not increase apoptosis. Comparison of consecutive harvests showed no difference in the numbers of CD34+ cells collected but found a significant decrease in apoptotic CD34+ cells through multiple collections. Analysis of annexin V binding in PBSC harvests is a simple flow cytometry technique which gives additional information on the status of CD34+ progenitor cells.
authors.affiliation: University of Edinburgh, John Hughes Bennett Laboratory, Western General Hospital, UK.
authors.forename: R S
authors.initials: RS
authors.lastname: Anthony
authors.forename: N D
authors.initials: ND
authors.lastname: McKelvie
authors.forename: A J
authors.initials: AJ
authors.lastname: Cunningham
authors.forename: J I
authors.initials: JI
authors.lastname: Craig
authors.forename: S Y
authors.initials: SY
authors.lastname: Rogers
authors.forename: A C
authors.initials: AC
authors.lastname: Parker
mesh_terms: D017304:Annexin A5; D018952:Antigens, CD34 / Q000097:blood; D018906:Antineoplastic Agents, Alkylating / Q000627:therapeutic use; D017209:Apoptosis; D001781:Blood Component Removal; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D003520:Cyclophosphamide / Q000627:therapeutic use; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate; D016179:Granulocyte Colony-Stimulating Factor / Q000627:therapeutic use; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D007938:Leukemia / Q000188:drug therapy / Q000503:physiopathology*; D008223:Lymphoma / Q000188:drug therapy / Q000503:physiopathology*
chemical_list: D017304:Annexin A5; D018952:Antigens, CD34; D018906:Antineoplastic Agents, Alkylating; D016179:Granulocyte Colony-Stimulating Factor; D003520:Cyclophosphamide; D016650:Fluorescein-5-isothiocyanate