Clinical-scale expansion of CD34+cord blood cells amplifies committed progenitors and rapid scid repopulation cells.
Umbilical cord blood (UCB) transplantation is associated with long periods of aplastic anaemia. This undesirable situation is due to the low cell dose available per unit of UCB and the immaturity of its progenitors. To overcome this, we present a cell culture strategy aimed at the expansion of the CD34+population and the generation of granulocyte lineage-committed progenitors. Two culture products were produced after either 6 or 14days of in vitro expansion, and their characteristics compared to non-expanded UCB CD34+controls in terms of phenotype, colony-forming activity and multilineage repopulation potential in NOD-scid IL2Rγnullmice. Both expanded cell products maintained rapid SCID repopulation activity similar to the non-expanded control, but 14-day cultured cells showed impaired long term SCID repopulation activity. The process was successfully scaled up to clinically relevant doses of 89×106CD34+cells committed to the granulocytic lineage and 3.9×109neutrophil precursors in different maturation stages. Cell yields and biological properties presented by the cell product obtained after 14days in culture were superior and therefore this is proposed as the preferred production setup in a new type of dual transplant strategy to reduce aplastic periods, producing a transient repopulation before the definitive engraftment of the non-cultured UCB unit. Importantly, human telomerase reverse transcriptase activity was undetectable, c-myc expression levels were low and no genetic abnormalities were found, as determined by G-banding karyotype, further confirming the safety of the expanded product.
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain.
Alba
A
Casamayor-Genescà
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain.
Arnau
A
Pla
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain.
Irene
I
Oliver-Vila
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain.
Noèlia
N
Pujals-Fonts
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain.
Sílvia
S
Marín-Gallén
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain.
Marta
M
Caminal
Immunology Department, Institut de Recerca Germans Trias i Pujol, Carretera Canyet s/n, Badalona, Spain.
Irma
I
Pujol-Autonell
Immunology Department, Institut de Recerca Germans Trias i Pujol, Carretera Canyet s/n, Badalona, Spain.
Jorge
J
Carrascal
Immunology Department, Institut de Recerca Germans Trias i Pujol, Carretera Canyet s/n, Badalona, Spain.
Marta
M
Vives-Pi
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain; Chair of Transfusion Medicine and Cellular and Tissue Therapies, Universitat Autònoma de Barcelona, Campus UAB, Cerdanyola del Vallès, 08193 Bellaterra, Spain.
Joan
J
Garcia
Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, 116, 08005 Barcelona, Spain. Electronic address: jvives@bst.cat.
Joaquim
J
Vives
D000741:Anemia, Aplastic / Q000097:blood / Q000209:etiology / Q000517:prevention & control; D000818:Animals; D018952:Antigens, CD34 / Q000097:blood; D001709:Biotechnology; D002454:Cell Differentiation; D019070:Cell Lineage; D003114:Colony-Forming Units Assay; D036101:Cord Blood Stem Cell Transplantation / Q000009:adverse effects / Q000379:methods*; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology; D006082:Graft Enhancement, Immunologic / Q000379:methods; D006098:Granulocytes / Q000166:cytology; D006801:Humans; D053631:Interleukin Receptor Common gamma Subunit / Q000172:deficiency / Q000235:genetics; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D009504:Neutrophils / Q000166:cytology
D018952:Antigens, CD34; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit
Aplasia; Bioreactor; CD34(+); Cell culture; Clinical scale; Hematopoietic stem cell transplantation; Umbilical cord blood