原创力文档antibody-directed lentiviral gene transduction for live-cell monitoring and selection of human ips and hes cellsantibody-directed慢病毒基因转导live-cell监测和选择人类ips和他的细胞.pdfVIP
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antibody-directed lentiviral gene transduction for live-cell monitoring and selection of human ips and hes cellsantibody-directed慢病毒基因转导live-cell监测和选择人类ips和他的细胞
Antibody-Directed Lentiviral Gene Transduction for Live-
Cell Monitoring and Selection of Human iPS and hES
Cells
3 2 1 2 1
Dai-tze Wu , Yasunari Seita , Xia Zhang , Chi-Wei Lu , Monica J. Roth *
1 Department of Biochemistry, University of Medicine and Dentistry of New Jersey – Robert Wood Johnson Medical School, Piscataway, New Jersey, United States of
America, 2 Deptartment of Ob/Gyn, University of Medicine and Dentistry of New Jersey – Robert Wood Johnson Medical School, Piscataway, New Jersey, United States of
America, 3 Department of Pharmacology, University of Medicine and Dentistry of New Jersey – Robert Wood Johnson Medical School, Piscataway, New Jersey, United
States of America
Abstract
The identification of stem cells within a mixed population of cells is a major hurdle for stem cell biology–in particular, in the
identification of induced pluripotent stem (iPS) cells during the reprogramming process. Based on the selective expression
of stem cell surface markers, a method to specifically infect stem cells through antibody-conjugated lentiviral particles has
been developed that can deliver both visual markers for live-cell imaging as well as selectable markers to enrich for iPS cells.
Antibodies recognizing SSEA4 and CD24 mediated the selective infection of the iPS cells over the parental human
fibroblasts, allowing for rapid expansion of these cells by puromycin selection. Adaptation of the vector allows for the
selective marking of human embryonic stem (hES) cells for their removal from a population of differentiated cells. This
method has the benefit that it not only identifies stem cells, but that specific genes, including positive and negative
selection markers, regulatory genes or miRN
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