1. Academic Validation
  2. A fluorescent chemical probe CDy9 selectively stains and enables the isolation of live naïve mouse embryonic stem cells

A fluorescent chemical probe CDy9 selectively stains and enables the isolation of live naïve mouse embryonic stem cells

  • Biomaterials. 2018 Oct;180:12-23. doi: 10.1016/j.biomaterials.2018.07.007.
Seung-Ju Cho 1 Keun-Tae Kim 1 Jong-Soo Kim 2 Ok-Seon Kwon 3 Young-Hyun Go 1 Nam-Young Kang 4 Haejeong Heo 5 Mi-Rang Kim 5 Dong Wook Han 6 Sung-Hwan Moon 2 Young-Tae Chang 7 Hyuk-Jin Cha 8
Affiliations

Affiliations

  • 1 Department of Life Sciences Sogang University, 35 Baeckbeom-ro, Mapo-gu, Seoul 04107, Republic of Korea.
  • 2 Department of Medicine, School of Medicine Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea.
  • 3 College of Pharmacy, Department of Pharmacy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea.
  • 4 Singapore Bioimaging Consortium, Agency for Science, Technology and Research, 11 Biopolis Way, Singapore 138667, Singapore.
  • 5 Personalized Genomic Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea.
  • 6 Department of Bioscience and Biotechnology, Bio-Organ Research Center, Konkuk University, Seoul, South Korea.
  • 7 Department of Chemistry, Pohang University of Science and Technology (POSTECH), Pohang 37673, Republic of Korea; Center for Self-assembly and Complexity, Institute for Basic Science (IBS), Pohang 37673, Republic of Korea. Electronic address: ytchang@postech.ac.kr.
  • 8 College of Pharmacy, Department of Pharmacy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea. Electronic address: hjcha93@snu.ac.kr.
Abstract

Human and mouse embryonic stem cells (ESCs) differ in terms of their pluripotency status, i.e., naïve vs. primed. This affects various biological properties and leads to several technical hurdles for future clinical applications, such as difficulties in chimera formation, single-cell passaging, and gene editing. In terms of generating functional human tissues and organs via mammalian interspecies chimerism, a fluorescent chemical probe that specifically labels naïve ESCs would help to isolate these cells and monitor their conversion. This study demonstrates that the fluorescent chemical probe compound of designation yellow 9 (CDy9) selectively stains naïve, but not primed, mouse ESCs (mESCs). CDy9 entered cells via Slc13a5, a highly expressed membrane transporter in naïve mESCs. Fluorescence-based cell sorting based on CDy9 staining successfully separated naïve mESCs from primed mESCs. Mice generated using CDy9+ cells isolated during the conversion of mouse epiblast stem cells into naïve mESCs exhibited coat color chimerism. Furthermore, CDy9 specifically stained cells in the inner cell mass of mouse embryos. These findings suggest that CDy9 is a useful tool to isolate functional naïve mESCs.

Keywords

Fluorescence probe; Inner cell mass; Live isolation; Naïve ESCs; Primed ESCs; Slc13a5.

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