AIM: We investigated the heterogeneity of tafluprost metabolism in primary human hepatocytes at a single-cell level by live single-cell mass spectrometry (MS). MATERIALS & METHODS: Picoliter volumes of cytoplasm were analyzed by nano-electrospray ionization MS in order to obtain single-cell metabolite profiles. The subcellular components of a single tafluprost-treated human hepatocyte were isolated and the single-cell metabolite profile was compared with those of traditional bulk hepatocyte analysis. RESULTS: In the bulk hepatocyte analysis, liquid chromatography-MS showed the averaged metabolism of tafluprost to tafluprost acid (TA) and β-oxidized metabolites. However, live single-cell MS showed that tafluprost metabolism varied among individual cells. In addition, there was significant variation in the quantities of TA and a major metabolite, dinor-TA, among cells, whereas there was no significant variation in 7-ethoxycoumarin metabolism. CONCLUSION: Thus, live single-cell MS successfully detected the heterogeneity of drug metabolism in individual living hepatocytes.
AIM: We investigated the heterogeneity of tafluprost metabolism in primary human hepatocytes at a single-cell level by live single-cell mass spectrometry (MS). MATERIALS & METHODS: Picoliter volumes of cytoplasm were analyzed by nano-electrospray ionization MS in order to obtain single-cell metabolite profiles. The subcellular components of a single tafluprost-treated human hepatocyte were isolated and the single-cell metabolite profile was compared with those of traditional bulk hepatocyte analysis. RESULTS: In the bulk hepatocyte analysis, liquid chromatography-MS showed the averaged metabolism of tafluprost to tafluprost acid (TA) and β-oxidized metabolites. However, live single-cell MS showed that tafluprost metabolism varied among individual cells. In addition, there was significant variation in the quantities of TA and a major metabolite, dinor-TA, among cells, whereas there was no significant variation in 7-ethoxycoumarin metabolism. CONCLUSION: Thus, live single-cell MS successfully detected the heterogeneity of drug metabolism in individual living hepatocytes.
Authors: Bo Yang; Nathan Heath Patterson; Tina Tsui; Richard M Caprioli; Jeremy L Norris Journal: J Am Soc Mass Spectrom Date: 2018-03-13 Impact factor: 3.109
Authors: Tom M J Evers; Mazène Hochane; Sander J Tans; Ron M A Heeren; Stefan Semrau; Peter Nemes; Alireza Mashaghi Journal: Anal Chem Date: 2019-10-08 Impact factor: 6.986