Live Cell Imaging: Differentiate ES/iPS from differentiated cells

Kyoto probe - structure and emmission-extinction

Fig 1: Structure and Fluorescent feature of KP-1 (λex – 515 nm; λem – 529 nm)

In a lot of experimental setups it would be eligible to be able to differentiate human ES and iPS cells from differentiated cells.

tebu-bio is pleased to announce their cooperation with Goryo Chemical, a company specialized in probes for Live Cell Imaging. Goryo Chemical recently launched the Kyoto probe 1 (KP-1) which exactly fulfills this need.

Kyoto probe 1 –  applicable in live cell imaging and flow cytometry

ES vs diff cells

Fig 2: Mechanism of Kyoto probe 1

Fig 1 shows the structure and the biophysical properties of the new Kyoto 1 probe. KP-1 is cell permeable and localizes in the mitochondria in the human pluripotent stem cells regardless of mitochondrial membrane potential. After the differentiation of the cells, KP-1 is eliminated from the cells by the ABC transporters which do not work in the pluripotent stem cells, and the cells become non-fluorescent as shown in Fig 2 (1).

Applications in Live Cell Imaging

In Fig 3 a number of live cell imaging results are shown to demonstrate the capacity of KP-1 to distinguish between stem cells and differentiated cells.

LCI experiements

Fig 2: (A、B) Staining iPS colony, formed on the feeder cells, with KP-1. (A: bright field image, B: fluorescent image). (C、F) Staining colony of partially differentiated human iPS cells with KP-1. (C: bright field image, F: fluorescent image). Surrounding region of the colony, in which cells were not differentiated, were specifically stained. (D、G) Staining a colony of human ES cells with KP-1. (D: bright field image, G: fluorescent image). (E、H) Dyeing colony of partially differentiated human ES cells with KP-1. (E: bright field image, H: fluorescent image). Differentiated cells were not stained.

Applications in Flow Cytometry

Besides staining human ES/iPS cells for Live Cell Imaging applications, KP-1 can also be used for Flow Cytometry applications as specified in Fig 4.

FACS application

Fig 4: FACS analysis of human iPS cells (Left panel) When the mixture of human iPS cells and feeder cells were analyzed, most SSEA-4, pluripotent stem cell marker, positive cells were also positive to KP-1. SSEA-4 is pluripotent stem cell. (Center and Right panel) 99.18% of human iPS cells were stained by KP-1.

Broad range of new Live Cell Imaging tools available

Goryo Chemical offers a lot more. Tools to conduct Acidic Organelle Imaging, imaging of galctosidase, β-glucosidase and β-glucuronidase… Furthermore they offer probes to measure Nitric oxide (NO), reactive oxgen species, Fe2+ and Zn2+, Hypoxia detection probes… you can find a complete overview here.

Interested in the new KP-1 probe or in any other tool for Live Cell Imaging? Please leave your comment in the form below.

Reference:

(1) Hirata et al., A Chemical Probe that Labels Human Pluripotent Stem Cells, Cell Reports 6, 6, p 1165–1174 (2014)

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Written by Ali El Baya, PhD
Ali el Bayâ is the Sales Manager at tebu-bio for the North of Europe.