Abstract
The study and application of human pluripotent stem cells (hPSCs) will be enhanced by the availability of well‐characterized monoclonal antibodies (mAbs) detecting cell‐surface epitopes. Here, we report generation of seven new mAbs that detect cell surface proteins present on live and fixed human ES cells (hESCs) and human iPS cells (hiPSCs), confirming our previous prediction that these proteins were present on the cell surface of hPSCs. The mAbs all show a high correlation with POU5F1 (OCT4) expression and other hPSC surface markers (TRA‐160 and SSEA‐4) in hPSC cultures and detect rare OCT4 positive cells in differentiated cell cultures. These mAbs are immunoreactive to cell surface protein epitopes on both primed and naive state hPSCs, providing useful research tools to investigate the cellular mechanisms underlying human pluripotency and states of cellular reprogramming. In addition, we report that subsets of the seven new mAbs are also immunoreactive to human bone marrow‐derived mesenchymal stem cells (MSCs), normal human breast subsets and both normal and tumorigenic colorectal cell populations. The mAbs reported here should accelerate the investigation of the nature of pluripotency, and enable development of robust cell separation and tracing technologies to enrich or deplete for hPSCs and other human stem and somatic cell types. Stem Cells 2017;35:626–640
Highlights
Human ESCs [1] and iPSCs [2, 3] have revolutionized the possibilities for cell-based regenerative therapies, but there are few in vitro diagnostic approaches that facilitate quality control of live human pluripotent stem cells and their derivatives
We report generation of seven new monoclonal antibodies (mAbs) that detect cell surface proteins present on live and fixed human ES cells and human iPS cells, confirming our previous prediction that these proteins were present on the cell surface of human pluripotent stem cells (hPSCs)
To generate tools for detecting cell-surface proteins on viable hPSCs that correlate with the presence of the pluripotencyassociated transcription factor OCT4 [50], we selected candidate genes that we identified from our fluorescence activated cell sorting (FACS)-based GCTM-2/CD9 immunotranscriptional profiling of hPSCs [25]
Summary
Human ESCs [1] and iPSCs [2, 3] have revolutionized the possibilities for cell-based regenerative therapies, but there are few in vitro diagnostic approaches that facilitate quality control of live human pluripotent stem cells (hPSCs) and their derivatives. Refinement of such approaches is an essential requirement for the safe and effective translation to diagnostic and therapeutic applications of hPSC-derived cell types [4, 5].
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