Diversity Oriented Fluorescence Library Approach for Stem Cell Probe Development

Abstract

Directed differentiation of stem cells and reprogramming of somatic cells into stem cells are the key issues in stem cell research and regenerative medicine. The most demanding requisites in the basic research and clinical applications of stem cells is to develop tools and methodologies for detecting and isolating specific type of stem cells at different stages of differentiation and reprogramming. Bioimaging which employs highly sophisticated imaging probes is becoming an emerging and rapidly growing field in biomedicine. Although stem cells have been visualized by various imaging techniques including fluorescence, luminescence, MRI, PET, and SPECT, the development of more specific and reliable imaging probes is an unmet need. Optical imaging techniques employing fluorescence have particular advantages in terms of detectability, efficiency and applicability in the bioimaging probe development. Using combinatorial chemistry, we have developed Diversity Oriented Fluorescence Library (DOFL) composed of intrinsically fluorescent small molecule collections. The power of DOFL approach has been demonstrated by the development of sensors and imaging probes for DNA, RNA, GTP, human serum albumin, glutathione, heparin, beta amyloid plaque and differentiated muscle cell. These successful results demonstrate that the DOFL can be applied, due to the unbiased structural diversity, to the screening of various analytes thus maximizing the chance of successful development of bioimaging probes. By screening a diversity oriented rosamine library, we developed the first fluorescent pluripotent stem cell probe CDy1, which also detects the cells undergoing reprogramming into induced pluripotent stem cells.