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Suspension Culture of Undifferentiated Human Embryonic and Induced Pluripotent Stem Cells
Authors:Michal Amit  Judith Chebath  Victoria Margulets  Ilana Laevsky  Yael Miropolsky  Kohava Shariki  Meital Peri  Idit Blais  Guy Slutsky  Michel Revel  Joseph Itskovitz-Eldor
Affiliation: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
2. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel
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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