Kyeong-Mo Kooa,1, Young-Hyun Goc,d,1, Seong-Min Kimd, Chang-Dae Kima, Jeong Tae Dob, Tae-Hyung Kima, Hyuk-Jin Chad
aSchool of Integrative Engineering, Chung-Ang University, Seoul, 06974, Republic of Korea
bDepartment of Stem Cell and Regenerative Biology, College of Animal Bioscience and Technology, Konkuk University, Seoul, Republic of Korea
cResearch Institute of Pharmaceutical Science, Seoul National University, Seoul, 08826, Republic of Korea
dCollege of Pharmacy, Seoul National University, Seoul, 08826, Republic of Korea
1These authors contributed equally.
Corresponding authors: Tae-Hyung Kim, Hyuk-Jin Cha
Pluripotent stem cells (PSCs) exist in naïve or primed states based on their origin. For in vitro culture, these PSCs require different supplements and growth factors. However, owing to their similar phenotypic features, identifying both cell types without harming cellular functions is challenging. This study reports an electrochemical method that enables simple, label-free, and non-destructive detection of naïve embryonic stem cells (ESCs) derived from mouse ESCs, based on the differences in cellular metabolism. Two major metabolic pathways to generate adenosine triphosphate (ATP)—glycolysis and oxidative phosphorylation (OXPHOS)—were blocked, and it was found that mitochondrial energy generation is the origin of the strong electrochemical signals of naïve ESCs. The number of ESCs is quantified when mixed with primed ESCs or converted from naïve–primed switchable metastable ESCs. The mouse PSCs derived from doxycycline-inducible mouse embryonic fibroblasts (MEFs) are also sensitively identified among other cell types such as unconverted MEFs and primed PSCs. The developed sensing platform operates in a non-invasive and label-free manner. Thus, it can be useful in the development of stem cell-derived therapeutics.