Induced Pluripotent Stem Cells
Abstract
Human pluripotent stem cells (hPSC) as typified by human embryonic stem cells (hESC) and recently by 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 robust and large scale reprogramming of somatic cells towards a pluripotent state need to be developed to leverage the potential of induced pluripotent stem cell technologies in drug discovery and regenerative medicine. Recently, HAMILTON and Global Cell Solutions™ developed a complete 3D cell culture system that streamlines the cell culture process for conventional cell lines and types. Here we present preliminary results on the use of this automated 3D culture system to reprogram human somatic cells in such a non-conventional but highly scalable culture system.
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
The generation of iPSC from a variety of differerent subjects (e.g. patients) is a cumbersome and expensive process. It will be important to develop high-throughput methods for reprogramming in semi- or fully automated systems. The GEM™ and BioLevitator™ is a system suitable for culturing somatic cells as well as fully reprogrammed iPSC . Protocols to reprogram somatic cells on this or any other 3D system still need to be developed and refined. |
Click the "Downloads and Links" tab for more details about the culture protocol and select the file titled Reprogramming Towards Induced Pluripotency in a 3D Culture System.
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