title: Functional studies of maturing myeloid cells during ex vivo expansion for treatment of aplasia: feasibility of ex vivo expansion from cryopreserved bone marrow cell samples.
abstract: Ex vivo expanded CD34+ progenitor cells from fresh or cryopreserved primate bone marrow, induced to granulocytic differentiation with growth factors, were investigated to determine whether myeloid cells produced in liquid cultures have the normal biologic functions needed for the treatment of patients with neutropenia following high-dose chemotherapy or therapeutic or accidental radiation exposure. Human and simian (baboons or macaques) CD34+ cells were cultured with granulocyte-colony stimulating factor (G-CSF), stem cell factor (SCF), interleukin-1 (IL-1), IL-3, and IL-6, and assessed at 14 days of culture for their capacity to respond to different functional tests. Immunostaining revealed that human ex vivo expanded cells contained myeloperoxydase (MPO, 82% +/- 8%) and lactoferrin (LF, 30% +/- 6%) in their granules. Maturation of cultured cells was associated with stimulated chemotactic responsiveness and respiratory burst activity (superoxide anion and hydrogen peroxide production) in expansions from human, baboon, and macaque CD34+ progenitor cells. Mature cells obtained from ex vivo expansion of selected cryopreserved human bone marrow CD34+ cells presented reduced but significant functional activities (chemotactic responsiveness and hydrogen peroxide production) when compared with human peripheral blood neutrophils. The validation of nonhuman primate ex vivo expansion systems may permit their use as models of irradiation. The feasibility of ex vivo expansion from cryopreserved bone marrow cell samples may offer considerable opportunity for banking bone marrow for autologous transfusion.
authors.affiliation: Institut de Protection et de Sûreté Nucléaire, Département de Protection de la santé de I'Homme et de Dosimétrie, Fontenay-aux-Roses, France.
authors.forename: T M
authors.initials: TM
authors.lastname: Neildez-Nguyen
authors.forename: J
authors.initials: J
authors.lastname: Vétillard
authors.forename: M
authors.initials: M
authors.lastname: Drouet
authors.forename: F
authors.initials: F
authors.lastname: Hérodin
authors.forename: N
authors.initials: N
authors.lastname: Brouard
authors.forename: J C
authors.initials: JC
authors.lastname: Mestries
authors.forename: D
authors.initials: D
authors.lastname: Thierry
mesh_terms: D000741:Anemia, Aplastic / Q000628:therapy*; D000818:Animals; D018952:Antigens, CD34 / Q000097:blood*; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D015925:Cryopreservation; D005240:Feasibility Studies; D006412:Hematopoietic Stem Cells / Q000276:immunology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007781:Lactoferrin / Q000097:blood; D008252:Macaca fascicularis; D009503:Neutropenia / Q000628:therapy; D010215:Papio; D009195:Peroxidase / Q000097:blood; D013481:Superoxides / Q000378:metabolism
chemical_list: D018952:Antigens, CD34; D013481:Superoxides; D006861:Hydrogen Peroxide; D009195:Peroxidase; D007781:Lactoferrin