Fluorescent Probe for Specific Staining of Human iPS/ES Cells
Kyoto Probe 1 (KP-1)
Fluorescence Spectra of KP-1
iPS Colony Formed on Feeder Cells; Stained with KP-1.
A: Bright-field microscopy, B: Fluorescence microscopy
KP-1 Selectivity Depends on the Activity of ABC Transporters in the Cell Membrane
KP-1 is cell permeable and its subcellular localization is in mitochondria. Only mitochondria of human pluripotent stem cells are stained by KP-1. The staining properties of KP-1 are independent of the mitochondrial membrane potential. After cell differentiation, KP-1 is eliminated from cells by ABC transporters which are not active in pluripotent stem cells, and cells become non-fluorescent. Expression levels of ABC transporters are markedly higher in differentiated cells than in human pluripotent stem cells. (Cell Rep. 2014).
C & F: Colony of partially differentiated human iPS cells stained with KP-1. The surrounding region of the colony, where cells were not differentiated, was specifically stained.
D & G: Colony of human ES cells stained with KP-1.
E & H: Colony of partially differentiated human ES cells stained with KP-1. Differentiated cells were not stained.
(C, D, E: Bright-field microscopy; F, G, H: Fluorescence microscopy).
Staining Results of KP-1 with Various Human Somatic Cells
After staining with KP-1 human iPS cells can be isolated from somatic cells by flow cytometry, except neuronal cells. These results indicate that KP-1 is useful for monitoring a wide range of differentiation processes of human pluripotent stem cells, except neurogenesis.
FACS Analysis of Human iPS Cells
Left panel: When a mixture of human iPS cells and feeder cells were analyzed, most SSEA-4 positive cells were also positive to KP-1. SSEA-4 is a well-known cell surface marker for pluripotent stem cells.
Center and right panel: 99.18% of human iPS cells were stained by KP-1.
- Kyoto Probe 1 (KP-1) distinguishes human iPS/ES cells from differentiated cells
- KP-1 selects for human iPS/ES cells due to ABC transporter expression
- For flow cytometry or live cell imaging
- Staining during cultivation of the cells
Fluorescence Characteristics of KP-1
- Kyoto Probe 1 [KP-1] – a small-molecule fluorescent compound – selectively labels human stem cells. Therefore, it is a highly appropriate tool for basic cell biology research and stem cell therapy. The selectivity results primarily from a distinct expression pattern of ABC transporters in human stem cells and from the transporter selectivity for KP-1. Expression of ABCB1 and ABCG2, both of which cause the efflux of KP-1, is repressed in human iPS/ES cells.
KP-1 Distinguishes Between Human iPSCs/ESCs and Differentiated Cells
Specific Staining of Human iPS/ES Cells by KP-1 - Basis for Selectivity
KP-1 is Suitable for Flow Cytometry and Live Cell Imaging
Isolation of Human iPS Cells by Flow Cytometry
- GORYO Fluorescent Probes Catalog (PDF)