期刊
BIOMEDICAL MICRODEVICES
卷 11, 期 1, 页码 87-94出版社
SPRINGER
DOI: 10.1007/s10544-008-9212-5
关键词
Micro-bioreactors; Perfusion system; Human embryonic stem cells; Cell culture in microsystems
资金
- Horowitz Foundation
- Israeli Cellome Network-ICN
- Aaron and Miriam Gutwirth Memorial Scholarship
The present study examines the use of automated periodic flow-stop perfusion systems for long-term culture of mammalian cells in a microchannel bioreactor. The method is used to culture Human Foreskin Fibroblasts (HFF) and Human Umbilical Vein Endothelial Cells (HUVEC) for long periods of time (> 7 d) in a microchannel (height 100 mu m). Design parameters, mass transport and shear stress issues are theoretically examined via numerical simulations. Cell growth and morphology are experimentally monitored and an enhanced growth rate was measured compared to constant perfusion micro-reactors and to traditional culture in Petri dishes. Moreover, we demonstrate the use of the method to co-culture undifferentiated colonies of human Embryonic Stem Cells (hESC) on HFF feeder cells in microchannels. The successful hESC-HFF co-culture in the microbioreactor is achieved due to two vital characteristics of the developed method-short temporal exposure to flow followed by long static incubation periods. The short pulsed exposure to shear enables shear sensitive cells (e.g., hESC) to withstand the medium renewal flow. The long static incubation period may enable secreted factors (e.g., feeder cells secreted factors) to accumulate locally. Thus the developed method may be suitable for long-term culture of sensitive multi-cellular complexes in microsystems.
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