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Merck
  • Antibody-directed lentiviral gene transduction for live-cell monitoring and selection of human iPS and hES cells.

Antibody-directed lentiviral gene transduction for live-cell monitoring and selection of human iPS and hES cells.

PloS one (2012-04-27)
Dai-tze Wu, Yasunari Seita, Xia Zhang, Chi-Wei Lu, Monica J Roth
摘要

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 miRNA can be delivered to the targeted stem cells. The ability to specifically target gene delivery to human pluripotent stem cells has broad applications in tissue engineering and stem cell therapies.

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TRAPeze RT端粒酶检测试剂盒, A highly sensitive in vitro assay for the fluorometric detection & real time quantification of telomerase activity in cells.