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Suspension Culture of Undifferentiated Human Embryonic and Induced Pluripotent Stem Cells

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Abstract

Alongside their contribution to research, human embryonic stem cells (hESC) may also prove valuable for cell-based therapies. Traditionally, these cells have been grown in adhesion culture either with or without feeder cells, allowing for their continuous growth as undifferentiated cells. However, to be applicable in therapy and industry they must be produced in a scalable and controlled process. Here we present for the first time a suspension culture system for undifferentiated hESC and induced pluripotent stem cells (iPSC), based on medium supplemented with the IL6RIL6 chimera (interleukin-6 receptor fused to interleukin-6), and basic fibroblast growth factor. Four hESC lines cultured in this system maintained all ESC features after 20 passages, including stable karyotype and pluripotency. Similar results were obtained when hESC were replaced with iPSC from two different cell lines. We demonstrate that the IL6RIL6 chimera supports the self-renewal and expansion of undifferentiated hESC and iPSC in suspension, and thus present another efficient system for large-scale propagation of undifferentiated pluripotent cells for clinical and translational applications.

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Acknowledgments

We thank Mrs. Hadas O’Neill for editing the manuscript. This research was supported by Technion Research and Development Foundation (TRDF). J.I.-E. holds the Sylvia and Stanley Shirvan Chair in Cell and Tissue Regeneration Research at the Technion—Israel Institute of Technology.

Conflict of interest

The authors declare no potential conflicts of interest.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Rambam Medical Center, Haifa, and the Sohnis and Forman Families Stem Cell Center, the Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, 31096, Israel

    Michal Amit, Victoria Margulets, Ilana Laevsky, Yael Miropolsky, Kohava Shariki, Meital Peri, Idit Blais & Joseph Itskovitz-Eldor

  2. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

    Judith Chebath, Guy Slutsky & Michel Revel

Authors
  1. Michal Amit
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  2. Judith Chebath
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  3. Victoria Margulets
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  4. Ilana Laevsky
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  5. Yael Miropolsky
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  6. Kohava Shariki
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  7. Meital Peri
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  8. Idit Blais
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  9. Guy Slutsky
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  10. Michel Revel
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  11. Joseph Itskovitz-Eldor
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Correspondence to Joseph Itskovitz-Eldor.

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Amit, M., Chebath, J., Margulets, V. et al. Suspension Culture of Undifferentiated Human Embryonic and Induced Pluripotent Stem Cells. Stem Cell Rev and Rep 6, 248–259 (2010). https://doi.org/10.1007/s12015-010-9149-y

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