Rapid expansion of human hematopoietic stem cells by automated control of inhibitory feedback signaling.
Clinical hematopoietic transplantation outcomes are strongly correlated with the numbers of cells infused. Anticipated novel therapeutic implementations of hematopoietic stem cells (HSCs) and their derivatives further increase interest in strategies to expand HSCs ex vivo. A fundamental limitation in all HSC-driven culture systems is the rapid generation of differentiating cells and their secreted inhibitory feedback signals. Herein we describe an integrated computational and experimental strategy that enables a tunable reduction in the global levels and impact of paracrine signaling factors in an automated closed-system process by employing a controlled fed-batch media dilution approach. Application of this system to human cord blood cells yielded a rapid (12-day) 11-fold increase of HSCs with self-renewing, multilineage repopulating ability. These results highlight the marked improvements that control of feedback signaling can offer primary stem cell culture and demonstrate a clinically relevant rapid and relatively low culture volume strategy for ex vivo HSC expansion.
Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON M5S 3E1, Canada.
Elizabeth
E
Csaszar
Daniel C
DC
Kirouac
Mei
M
Yu
WeiJia
W
Wang
Wenlian
W
Qiao
Michael P
MP
Cooke
Anthony E
AE
Boitano
Caryn
C
Ito
Peter W
PW
Zandstra
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D003198:Computer Simulation; D003470:Culture Media / Q000378:metabolism; D025461:Feedback, Physiological; D005312:Fetal Blood / Q000166:cytology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D019899:Paracrine Communication
D003470:Culture Media