Undifferentiated hESC
Abstract
Human pluripotent stem cells (hPSC) as typified by human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) with their self-renewal and differentiation properties
hold great promise for understanding and treating human disease. Many technical hurdles, including the need for reproducible large scale expansion and differentiation protocols, must be addressed
to apply stem cell technologies for drug discovery and regenerative medicine. Recently, many new hESC/hiPSC culture methods have been developed that are to various degrees feeder-free and
completely define. However scaling up and automating the hESC/hiPSC culture process for commercial applications continues to impede adoption of hESC/hiPSC. Recently, the Hamilton Company and Global Cell Solutions developed a novel alginate microcarrier cell culture system that streamlines the cell culture
process and enables efficient scale up of hESCs. Here we present results on the use of this 3D culture system to propagate undifferentiated hPSC.
Introduction
Human pluripotent stem cell (hPSC) culture has improved significantly over the last 3 years:
• Once highly time-consuming and complex.
• Highly trained and experienced staff was and (is!) absolutely necessary for successful
hPSC culture
• Irradiated MEFs were required: quality and regulatory issues
• Time-consuming passaging procedures for relatively few cells
• Now development of commercially available, partially or completely defined feeder free
culture systems: mTeSR1 (Stemcell), StemPro SFM (Invitrogen), NutriStem™ XF/FF
(Stemgent)
Still several problems to solve:
• modern hPSC culture reagents are expensive,
• hPSC culture is still technically more demanding than with other cell types,
• large-scale culture (e.g. for proteomics or ultra deep sequencing experiments) requires
significant staff resources
HAMILTON Bonaduz AG & Global Cell Solutions ™ are developing a complete 3-D cell
culture system that streamlines the cell culture process for conventional cell lines and types:
• GEMs™ (Global Eucaryotic Microcarriers™) are magnetic alginate microcarrier, which
can be coated with extracellular matrices
• The BioLevitator R3 supports the complete cell culture process from inoculation to
harvesting in as a bench-top incubator that homogenizes continuously the GEMs. |
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Conclusion
Culture of hESC in a highly scalable 3D cell culture system holds great promise.
It will be important to develop very well characterized protocols which are directed at the culture of epigentically and karyotypically normal hESC.
The GEM™ and BioLevitator™ is a system suitable for culturing hESC. Yet, we have to be cautious, since certain culture procedures, which are currently
regarded as acceptable practice in the field, may cause significant epigentic and genetic alterations in long term cultures of undifferentiated pluripotent stem cells .
Hence, protocols to culture hESC on this or any other 3D system still need to be further developed and refined. |
Click the "Downloads and Links" tab for more details about the culture protocol and select the file titled Automated 3D Culture of Undifferentiated hESC.
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