Measuring ATP Concentration in a Small Number of Murine Hematopoietic Stem Cells
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The metabolism of quiescent adult stem cells differs from the metabolism of differentiated cells. The metabolic processes are tightly regulated and their alterations disturb function of stem cells. One of the indicators of metabolic status of cells is the ATP level. While the method of measuring the ATP levels has been known for many years, estimating ATP levels in small population of defined stem cells isolated directly from the tissue has remained challenging. Here, we show our method of measuring the ATP levels in hematopoietic stem cells sorted from murine bone marrow. We used magnetic sorting as well as cell sorter and adopted the commonly used bioluminescence-based detection kits in described protocol. Our strategy allows to measure ATP levels in 1000 highly purified HSC.
KeywordsHematopoietic stem cells ATP Metabolism Cell sorting
The research was supported by the following grants: HARMONIA (NCN 2015/18/M/NZ3/00387) granted to Alicja Jozkowicz by Polish National Science Centre and PRELUDIUM (NCN 2013/11/N/NZ3/00956) granted to Krzysztof Szade by Polish National Science Centre. The Faculty of Biochemistry, Biophysics, and Biotechnology of Jagiellonian University is a partner of the Leading National Research Centre (KNOW) supported by the Ministry of Science and Higher Education.