Suspension culture of undifferentiated human embryonic and induced pluripotent stem cells.
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.
Amit
Michal
M
Department of Obstetrics and Gynecology, Rambam Medical Center, Haifa, and Sohnis and Forman Families Stem Cell Center, Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, 31096, Israel.
Chebath
Judith
J
Margulets
Victoria
V
Laevsky
Ilana
I
Miropolsky
Yael
Y
Shariki
Kohava
K
Peri
Meital
M
Blais
Idit
I
Slutsky
Guy
G
Revel
Michel
M
Itskovitz-Eldor
Joseph
J
D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007621:Karyotyping; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology
C508600:IL6 protein, human; D015850:Interleukin-6; D011993:Recombinant Fusion Proteins