Abstract
Semiconductor quantum dots (QDs) hold increasing potential for cellular imaging both in vitro and in vivo. In this report, we aimed to study imaging of human embryonic stem (ES) cells labeled with quantum dots (QDs), and to evaluate the viability and pluripotency of human ES cells labeled with QDs. We used the Tat-QDs to label human ES cells, evaluated the cytoactivity of human ES cells labeled with QDs by CCK8 assay and Flow cytometer, and verified the pluripotency of human ES cells labeled with QDs by differentiated the human ES cells into hematopoietic cells and neural cells. The result illustrated that human embryonic stem (ES) cells were labeled with QDs and intracellular QD number was associated with the dose of QDs. Human ES cell viability, proliferation, and pluripotency were not adversely affected by QDs compared with non-labeled control cells. In summary, this is the first report showing the QDs labeled hESCs could be differentiated into hematopoietic cells and neural cells. These results provide a promising tool for imaging stem cell therapy noninvasively in vivo.
Highlights
Stem cell therapy holds promise for treatment of intractable conditions such as Parkinson's disease, ischemic heart disease, diabetes, and degenerative joint diseases [1,2,3,4]
Mouse Embryonic Fibroblast (MEF) cells were maintained in Dulbecco’s modified eagle medium (DMEM, GIBCO) containing 10% fetal bovine serum (FBS, GIBCO) and 1.0% penicillin and streptomycin (GIBCO). human Embryo Stem cell (hESC) were maintained in hESC completed medium which made up with DMEM/F12 (GIBCO) supplemented with knockout SR (GIBCO), bFGF (Invitrogen), NEAA (GIBCO), L-glutamine (GIBCO) and β-mercaptoethanol (GIBCO). hESCs were cultured on MEF feeder cells dish which were coated with Matrigel(BD) in advance according to the standard human embryonic stem cell culture protocol [17]
We carefully examined the effect of TatQDs on human ES cells by CCK8 assay
Summary
Stem cell therapy holds promise for treatment of intractable conditions such as Parkinson's disease, ischemic heart disease, diabetes, and degenerative joint diseases [1,2,3,4]. With QDs' many advantages over traditional organic dyes, QDs may provide an excellent tool for imaging stem cell therapy. Quantum dots (QDs) are nano-scale fluorescent semiconductors that are increasingly used as label tools in biological research. QDs have recently been utilized as fluorescent tags for label experiments in vivo and in vitro. They can be tailored, through control of size, composition, and shape, to provide broad spectral coverage with symmetric narrow emission profiles. QDs could be used as multicolor imaging applications and the tracking of live cells [6,7]. Most QD applications have utilized non-mammalian or cancer cells with only a few studies examining deleterious effects of QDs in human ES cells
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