title: Serial transplantations in nonobese diabetic/severe combined immunodeficiency mice of transduced human CD34+ cord blood cells: efficient oncoretroviral gene transfer and ex vivo expansion under serum-free conditions.
abstract: Stable oncoretroviral gene transfer into hematopoietic stem cells (HSCs) provides permanent genetic disease correction. It is crucial to transplant enough transduced HSCs to compete with and replace the defective host hemopoiesis. To increase the number of transduced cells, the role of ex vivo expansion was investigated. For a possible clinical application, all experiments were carried out in serum-free media. A low-affinity nerve growth factor receptor (LNGFR) pseudotyped murine retroviral vector was used to transduce cord blood CD34(+) cells, which were then expanded ex vivo. These cells engrafted up to three generations of serially transplanted nonobese diabetic/severe combined immunodeficiency mice: 54.26% +/- 5.59%, 19.05% +/- 2.01%, and 6.15% +/- 5.16% CD45(+) cells from primary, secondary, and tertiary recipient bone marrow, respectively, were LNGFR(+). Repopulation in secondary and tertiary recipients indicates stability of transgene expression and long-term self-renewal potential of transduced HSCs, suggesting that retroviral gene transfer into HSCs, followed by ex vivo expansion, could facilitate long-term engraftment of genetically modified HSCs.
authors.affiliation: Institute for Cancer Research and Treatment, Laboratory of Clinical Oncology, 10060 Candiolo, Torino, Italy.
authors.forename: Loretta
authors.initials: L
authors.lastname: Gammaitoni
authors.forename: Simona
authors.initials: S
authors.lastname: Lucchi
authors.forename: Stefania
authors.initials: S
authors.lastname: Bruno
authors.forename: Melania
authors.initials: M
authors.lastname: Tesio
authors.forename: Monica
authors.initials: M
authors.lastname: Gunetti
authors.forename: Ymera
authors.initials: Y
authors.lastname: Pignochino
authors.forename: Giorgia
authors.initials: G
authors.lastname: Migliardi
authors.forename: Lorenza
authors.initials: L
authors.lastname: Lazzari
authors.forename: Massimo
authors.initials: M
authors.lastname: Aglietta
authors.forename: Paolo
authors.initials: P
authors.lastname: Rebulla
authors.forename: Wanda
authors.initials: W
authors.lastname: Piacibello
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis*; D049109:Cell Proliferation; D002478:Cells, Cultured; D036101:Cord Blood Stem Cell Transplantation; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology / Q000637:transplantation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D012190:Retroviridae / Q000235:genetics*; D016511:Severe Combined Immunodeficiency / Q000235:genetics / Q000628:therapy*; D019076:Transgenes; D014183:Transplantation, Heterologous
chemical_list: D018952:Antigens, CD34; D016895:Culture Media, Serum-Free