Abstract
While human pluripotent stem cells are attractive sources for cell-replacement therapies, a major concern remains regarding their tumorigenic potential. Thus, safety assessment of human pluripotent stem cell-based products in terms of tumorigenicity is critical. Previously we have identified a pluripotent stem cell-specific lectin probe rBC2LCN recognizing hyperglycosylated podocalyxin as a cell surface ligand. Here we demonstrate that hyperglycosylated podocalyxin is secreted from human pluripotent stem cells into cell culture supernatants. We establish a sandwich assay system, named the GlycoStem test, targeting the soluble hyperglycosylated podocalyxin using rBC2LCN. The GlycoStem test is sufficiently sensitive and quantitative to detect residual human pluripotent stem cells. This work provides a proof of concept for the noninvasive and quantitative detection of tumorigenic human pluripotent stem cells using cell culture supernatants. The developed method should increase the safety of human pluripotent stem cell-based cell therapies.
Highlights
While human pluripotent stem cells are attractive sources for cell-replacement therapies, a major concern remains regarding their tumorigenic potential
In 2012, clinical trials have been conducted with retinal pigment epithelial (RPE) cells derived from hESCs to treat patients with dry age-related macular degeneration and Stargart’s macular dystrophy[3]
A major advantage of the developed sandwich assay system is that it requires cell culture supernatants, but not cells, for quantification of residual tumorigenic Human pluripotent stem cells (hPSCs), so that invaluable cells used for transplantation are not consumed
Summary
While human pluripotent stem cells are attractive sources for cell-replacement therapies, a major concern remains regarding their tumorigenic potential. Safety assessment of human pluripotent stem cell-based products in terms of tumorigenicity is critical. We have identified a pluripotent stem cell-specific lectin probe rBC2LCN recognizing hyperglycosylated podocalyxin as a cell surface ligand. This work provides a proof of concept for the noninvasive and quantitative detection of tumorigenic human pluripotent stem cells using cell culture supernatants. An in vivo teratoma formation assay using severe combined immunodeficiency (SCID) mice provides a straightforward means to assess the existence of tumorigenic stem cells in a cell population All of these currently available methods necessitate the use of a significant number (.104) of invaluable cells. This work provides a novel concept for the use of cell culture supernatants for the safety assessments of stem cell-based products
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