Monoclonal antibody K312-based depletion of pluripotent cells from differentiated stem cell progeny prevents teratoma formation
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Human pluripotent stem cells (PSCs) have been utilized as apromising source in regenerative medicine. However, the riskof teratoma formation that comes with residual undifferentiatedPSCs in differentiated cell populations is most concerning inthe clinical use of PSC derivatives. Here, we report that a monoclonalantibody (mAb) targeting PSCs could distinguish undifferentiatedPSCs, with potential teratoma-forming activity, fromdifferentiated PSC progeny. A panel of hybridomas generatedfrom mouse immunization with H9 human embryonic stem cells(hESCs) was screened for ESC-specific binding using flow cytometry. A novel mAb, K312, was selected considering its highstem cell-binding activity, and this mAb could bind to severalhuman induced pluripotent stem cells and PSC lines. Cell-bindingactivity of K312 was markedly decreased as hESCs weredifferentiated into embryoid bodies or by retinoic acid treatment. In addition, a cell population negatively isolated from undifferentiatedor differentiated H9 hESCs via K312 targeting showeda significantly reduced expression of pluripotency markers, includingOct4 and Nanog. Furthermore, K312-based depletionof pluripotent cells from differentiated PSC progeny completelyprevented teratoma formation. Therefore, our findings suggestthat K312 is utilizable in improving stem cell transplantationsafety by specifically distinguishing residual undifferentiated PSCs.
Abstract
Human pluripotent stem cells (PSCs) have been utilized as apromising source in regenerative medicine. However, the riskof teratoma formation that comes with residual undifferentiatedPSCs in differentiated cell populations is most concerning inthe clinical use of PSC derivatives. Here, we report that a monoclonalantibody (mAb) targeting PSCs could distinguish undifferentiatedPSCs, with potential teratoma-forming activity, fromdifferentiated PSC progeny. A panel of hybridomas generatedfrom mouse immunization with H9 human embryonic stem cells(hESCs) was screened for ESC-specific binding using flow cytometry. A novel mAb, K312, was selected considering its highstem cell-binding activity, and this mAb could bind to severalhuman induced pluripotent stem cells and PSC lines. Cell-bindingactivity of K312 was markedly decreased as hESCs weredifferentiated into embryoid bodies or by retinoic acid treatment. In addition, a cell population negatively isolated from undifferentiatedor differentiated H9 hESCs via K312 targeting showeda significantly reduced expression of pluripotency markers, includingOct4 and Nanog. Furthermore, K312-based depletionof pluripotent cells from differentiated PSC progeny completelyprevented teratoma formation. Therefore, our findings suggestthat K312 is utilizable in improving stem cell transplantationsafety by specifically distinguishing residual undifferentiated PSCs.
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