Abstract
We report that a single growth factor, NM23-H1, enables serial passaging of both human ES and iPS cells in the absence of feeder cells, their conditioned media or bFGF in a fully defined xeno-free media on a novel defined, xeno-free surface. Stem cells cultured in this system show a gene expression pattern indicative of a more “naïve” state than stem cells grown in bFGF-based media. NM23-H1 and MUC1* growth factor receptor cooperate to control stem cell self-replication. By manipulating the multimerization state of NM23-H1, we override the stem cell's inherent programming that turns off pluripotency and trick the cells into continuously replicating as pluripotent stem cells. Dimeric NM23-H1 binds to and dimerizes the extra cellular domain of the MUC1* transmembrane receptor which stimulates growth and promotes pluripotency. Inhibition of the NM23-H1/MUC1* interaction accelerates differentiation and causes a spike in miR-145 expression which signals a cell's exit from pluripotency.
Highlights
Both embryonic stem (ES) and induced pluripotent stem cells hold great promise for the treatment of a wide variety of acquired or hereditary diseases [1,2]
In an earlier study [41], we reported that a cleaved form of the MUC1 transmembrane receptor, called MUC1*, that had previously only been detected on cancer cells [42] is expressed on undifferentiated human embryonic stem cells and mediates their growth in an bFGF-independent manner
Their conditioned media, are widely used for the growth of ES and iPS cells, we investigated whether feeder cells were merely providing stem cells with NM23-H1.NM23-H1 was immuno-depleted from fibroblast feeder cell conditioned media (Fig. 1a, b) tested for the ability to support stem cell growth
Summary
Both embryonic stem (ES) and induced pluripotent stem (iPS) cells hold great promise for the treatment of a wide variety of acquired or hereditary diseases [1,2]. FDA and European guidelines for human stem cell therapies require some version of Good Manufacturing Practice (GMP) for quality assurance and patient safety. Most protocols used today involve a supporting layer of fibroblast feeder cells [7,8], their conditioned media [9] or Matrigel [10,11]. These are complex mixtures of poorly characterized components that vary greatly from batch to batch, and cannot be made GMPcompliant
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