title: Ex vivo expansion of autologous PB CD34+ cells provides a purging effect in children with neuroblastoma.
abstract: Peripheral blood CD34+ cell samples from eight children with advanced neuroblastoma and from 10 healthy adult donors were seeded at 5 x 10(4) cells/ml in stroma-free, serum-free medium with FL, SCF, MGDF (100 ng/ml each), G-CSF, IL6 (10 ng/ml each) and IL3 (5 ng/ml), and incubated for 10 days. The levels of expansion of PBCD34+ cells observed in neuroblastoma patients, with up to 214-fold expansion for total nucleated cells, 39-fold for CD34+ cells, 79-fold for CFU-GM and nine-fold for LTC-IC were identical to those obtained with PBCD34+ cells of healthy donors (P>/=0.5). All samples from patients with neuroblastoma and five donor's PBCD34+ cell samples contaminated with IMR-32 neuroblasts, were screened for the number of tyrosine hydroxylase (TH) mRNA transcript using LightCycler software. In all samples, progressive 1.9-4.4 log decreases in the number of TH transcripts were observed between days 0 and 10 of expansion. Our results show that in extensively pretreated children with neuroblastoma, the culture conditions that were effective for BM and CB cell expansion can generate an expansion of PBCD34+ cells and provide a purge of tumour cells.
authors.affiliation: Unité Bioclinique de Thérapie Cellulaire, Service de Pédiatrie B, Hôtel Dieu, C.H.U., B.P.69, 11, Boulevard Léon Malfreyt, Clermont-Ferrand 63003, France. jkanold@chu-clermontferrand.fr
authors.forename: J
authors.initials: J
authors.lastname: Kanold
authors.forename: P
authors.initials: P
authors.lastname: Halle
authors.forename: A
authors.initials: A
authors.lastname: Tchirkov
authors.forename: M
authors.initials: M
authors.lastname: Berger
authors.forename: M C
authors.initials: MC
authors.lastname: Giarratana
authors.forename: L
authors.initials: L
authors.lastname: Kobari
authors.forename: N
authors.initials: N
authors.lastname: Boiret
authors.forename: C
authors.initials: C
authors.lastname: Paillard
authors.forename: F
authors.initials: F
authors.lastname: Deméocq
authors.forename: L
authors.initials: L
authors.lastname: Douay
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002648:Child; D002675:Child, Preschool; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007223:Infant; D018365:Neoplasm, Residual / Q000175:diagnosis; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D009447:Neuroblastoma / Q000473:pathology / Q000628:therapy*; D036102:Peripheral Blood Stem Cell Transplantation / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis; D012334:RNA, Neoplasm / Q000032:analysis; D014182:Transplantation, Autologous; D014184:Transplantation, Homologous; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
chemical_list: D018952:Antigens, CD34; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D014446:Tyrosine 3-Monooxygenase