Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 94, Issue 5, Pages 938-948Publisher
WILEY
DOI: 10.1002/bit.20919
Keywords
human embryonic stem cell; hematopoiesis; embryoid body; bioreactor
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Funding
- NHLBI NIH HHS [R01 HL077923, HL 72000, R01 HL077923-01A2] Funding Source: Medline
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Human embryonic stem cells (hESCs) represent an important resource for novel cell-based regenerative medical therapies. hESCs are known to differentiate into mature cells of defined lineages through the formation of embryoid bodies (EBs) which are amenable to suspension culture for several weeks. However, EBs derived from hESCs in standard static cultures are typically non-homogeneous, leading to inefficient cellular development. Here, we systematically compare the formation, growth, and differentiation capabilities of hESC-derived EBs in stirred and static suspension cultures. A 15-fold expansion in total number of EB-derived cells cultured for 21 days in a stirred flask was observed, compared to a fourfold expansion in static (non-stirred) cultures. Additionally, stirred vessel mediated cultures have a more homogeneous EB morphology and size. Importantly, the EBs cultivated in spinner flasks retained comparable ability to produce hernatopoietic progenitor cells as those grown in static culture. These results demonstrate the decoupling between EB cultivation method and EB-derived cells' ability to form hematopoietic progenitors, and will allow for improved production of scalable quantities of hematopoietic cells or other differentiated cell lineages from hESCs in a controlled environment. (c) 2006 Wiley Periodicals, Inc.
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