Diversity Oriented Fluorescence Library Approach for Stem Cell Probe Development

  • Seong-Wook Yun
  • Nam-Young Kang
  • Young-Tae Chang
Part of the Stem Cells and Cancer Stem Cells book series (STEM, volume 3)


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.


DOFL CDy1 Superparamagnetic iron oxide SPECT PET Luminescence 


  1. Ahn YH, Lee JS, Chang YT (2007) Combinatorial rosamine library and application to in vivo glutathione probe. J Am Chem Soc 129:4510–4511PubMedCrossRefGoogle Scholar
  2. Alieva NO, Konzen KA, Field SF, Meleshkevitch EA, Hunt ME, Beltran-Ramirez V, Miller DJ, Wiedenmann J, Salih A, Matz MV (2008) Diversity and evolution of coral fluorescent proteins. PLoS One 3:e2680PubMedCrossRefGoogle Scholar
  3. Budde MD, Frank JA (2009) Magnetic tagging of therapeutic cells for MRI. J Nucl Med 50:171–174PubMedCrossRefGoogle Scholar
  4. Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805PubMedCrossRefGoogle Scholar
  5. Challen GA, Boles N, Lin KK, Goodell MA (2009) Mouse hematopoietic stem cell identification and analysis. Cytometry A 75:14–24PubMedGoogle Scholar
  6. Chan EM, Ratanasirintrawoot S, Park IH, Manos PD, Loh YH, Huo H, Miller JD, Hartung O, Rho J, Ince TA, Daley GQ, Schlaeger TM (2009) Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells. Nat Biotechnol 27:1033–1037PubMedCrossRefGoogle Scholar
  7. Chia NY, Chan YS, Feng B, Lu X, Orlov YL, Moreau D, Kumar P, Yang L, Jiang J, Lau MS, Huss M, Soh BS, Kraus P, LiP, Lufkin T, Lim B, Clarke ND, Bard F, Ng HH (2010) A genome-wide RNAi screen reveals determinants of human embryonic stem cell identity. Nature 468: 316–320PubMedCrossRefGoogle Scholar
  8. Cohen DE, Melton D (2011) Turning straw into gold: directing cell fate for regenerative medicine. Nat Rev Genet 12:243–252PubMedCrossRefGoogle Scholar
  9. Dothager RS, Flentie K, Moss B, Pan MH, Kesarwala A, Piwnica-Worms D (2009) Advances in bioluminescence imaging of live animal models. Curr Opin Biotechnol 20:45–53PubMedCrossRefGoogle Scholar
  10. Gilad AA, Winnard PT Jr, van Zijl PC, Bulte JW (2007) Developing MR reporter genes: promises and pitfalls. NMR Biomed 20:275–290PubMedCrossRefGoogle Scholar
  11. Hawley TS, Telford WG, Hawley RG (2001) “Rainbow” reporters for multispectral marking and lineage analysis of hematopoietic stem cells. Stem Cells 19:118–124PubMedCrossRefGoogle Scholar
  12. Hayashi H, Miyata H (1994) Fluorescence imaging of intracellular Ca2+. J Pharmacol Toxicol Methods 31:1–10PubMedCrossRefGoogle Scholar
  13. Huangfu D, Osafune K, Maehr R, Guo W, Eijkelenboom A, Chen S, Muhlestein W, Melton DA (2008) Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat Biotechnol 26:1269–1275PubMedCrossRefGoogle Scholar
  14. Im CN, Kang NY, Ha HH, Bi X, Lee JJ, Park SJ, Lee SY, Vendrell M, Kim YK, Lee JS, Li J, Ahn YH, Feng B, Ng HH, Yun SW, Chang YT (2010) A fluorescent rosamine compound selectively stains pluripotent stem cells. Angew Chem Int Ed Engl 49:7497–7500PubMedCrossRefGoogle Scholar
  15. Jiang H, Cheng Z, Tian M, Zhang H (2011) In vivo imaging of embryonic stem cell therapy. Eur J Nucl Med Mol Imaging 38:774–784PubMedCrossRefGoogle Scholar
  16. Kang JH, Chung JK (2008) Molecular-genetic imaging based on reporter gene expression. J Nucl Med 49(Suppl 2):164S–179SPubMedCrossRefGoogle Scholar
  17. Kennedy JP, Williams L, Bridges TM, Daniels RN, Weaver D, Lindsley CW (2008) Application of combinatorial chemistry science on modern drug discovery. J Comb Chem 10:345–354PubMedCrossRefGoogle Scholar
  18. Kim YK, Ha HH, Lee JS, Bi X, Ahn YH, Hajar S, Lee JJ, Chang YT (2010) Control of muscle differentiation by a mitochondria-targeted fluorophore. J Am Chem Soc 132:576–579PubMedCrossRefGoogle Scholar
  19. Kim YK, Lee JS, Bi X, Ha HH, Ng SH, Ahn YH, Lee JJ, Wagner BK, Clemons PA, Chang YT (2011) The binding of fluorophores to proteins depends on the cellular environment. Angew Chem Int Ed Engl 50:2761–2763PubMedCrossRefGoogle Scholar
  20. Kolle G, Ho M, Zhou Q, Chy HS, Krishnan K, Cloonan N, Bertoncello I, Laslett AL, Grimmond SM (2009) Identification of human embryonic stem cell surface markers by combined membrane-polysome translation state array analysis and immunotranscriptional profiling. Stem Cells 27:2446–2456PubMedCrossRefGoogle Scholar
  21. Lee S, Park K, Kim K, Choi K, Kwon IC (2008) Activatable imaging probes with amplified fluorescent signals. Chem Commun (Camb) 4250–4260Google Scholar
  22. Lee JS, Kim YK, Vendrell M, Chang YT (2009) Diversity-oriented fluorescence library approach for the discovery of sensors and probes. Mol Biosyst 5:411–421PubMedCrossRefGoogle Scholar
  23. Marshall IC, Owen DE, McNulty S (2005) Measuring ca(2+) changes in multiwell format using the fluorometric imaging plate reader. Methods Mol Biol 312:125–131PubMedGoogle Scholar
  24. Nagano K, Yoshida Y, Isobe T (2008) Cell surface biomarkers of embryonic stem cells. Proteomics 8:4025–4035PubMedCrossRefGoogle Scholar
  25. Paredes RM, Etzler JC, Watts LT, Zheng W, Lechleiter JD (2008) Chemical calcium indicators. Methods 46:143–151PubMedCrossRefGoogle Scholar
  26. Plath K, Lowry WE (2011) Progress in understanding reprogramming to the induced pluripotent state. Nat Rev Genet 12:253–265PubMedCrossRefGoogle Scholar
  27. Prasher DC, Eckenrode VK, Ward WW, Prendergast FG, Cormier MJ (1992) Primary structure of the Aequorea victoria green-fluorescent protein. Gene 111:229–233PubMedCrossRefGoogle Scholar
  28. Sacco A, Doyonnas R, Kraft P, Vitorovic S, Blau HM (2008) Self-renewal and expansion of single transplanted muscle stem cells. Nature 456:502–506PubMedCrossRefGoogle Scholar
  29. Vendrell M, Lee JS, Chang YT (2010) Diversity-oriented fluorescence library approaches for probe discovery and development. Curr Opin Chem Biol 14:383–389PubMedCrossRefGoogle Scholar
  30. Welling MM, Duijvestein M, Signore A, van der Weerd L (2011) In vivo biodistribution of stem cells using molecular nuclear medicine imaging. J Cell Physiol 226:1444–1452PubMedCrossRefGoogle Scholar
  31. Xie X, Chan KS, Cao F, Huang M, Li Z, Lee A, Weissman IL, Wu JC (2009) Imaging of STAT3 signaling pathway during mouse embryonic stem cell differentiation. Stem Cells Dev 18:205–214PubMedCrossRefGoogle Scholar
  32. Zijlmans JM, Visser JW, Kleiverda K, Kluin PM, Willemze R, Fibbe WE (1995) Modification of rhodamine staining allows identification of hematopoietic stem cells with preferential short-term or long-term bone marrow-repopulating ability. Proc Natl Acad Sci USA 92:8901–8905PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Seong-Wook Yun
    • 1
  • Nam-Young Kang
    • 1
  • Young-Tae Chang
    • 2
    • 3
  1. 1.Laboratory of Bioimaging Probe Development, Singapore Bioimaging ConsortiumAgency for Science, Technology and Research (A* STAR)SingaporeSingapore
  2. 2.NUS MedChem Program of Life Sciences Institute, National University of SingaporeSingaporeSingapore
  3. 3.Singapore Bioimaging ConsortiumAgency for Science, Technology and Research (A* STAR)SingaporeSingapore

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