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Fig. 3 | Journal of Translational Medicine

Fig. 3

From: Mesenchymal stromal cells derived from cervical cancer produce high amounts of adenosine to suppress cytotoxic T lymphocyte functions

Fig. 3

Hydrolytic activity of CD39 and CD73 ectonucleotidases expressed in MSCs. A total of 1 × 105 CEMs derived from NCx-MSCs (n = 5) and CeCa-MSCs (n = 5) were cultured at 37 °C with 5 mM adenine nucleotides (ATP, ADP or AMP) in the presence or absence of POM-1 (specific CD39 inhibitor) or APCP (specific CD73 inhibitor). a Adenosine produced by the hydrolysis of nucleotides was analyzed by thin layer chromatography (TLC). The ATP, ADP and AMP hydrolysis products (marked with arrows) at the end of MSC culture with different nucleotides are shown. ATP, ADP, AMP, inosine (Ino) and synthetic adenosine were used as markers. b Adenosine contained in supernatant samples was quantified every 60 min by ultra-performance liquid chromatography (UPLC), using standard concentrations of synthetic Ado (upper). A representative linear range between concentration and histogram integral area for Ado is shown (lower). c The concentration of Ado produced by the hydrolysis of ATP (upper), ADP (middle) and AMP (lower) during the period of MSC culture with the respective nucleotides is shown. Asterisk indicates significant (P < 0.001) differences compared to NCx-MSCs. d The concentrations of Ado produced by the hydrolysis of adenine nucleotides (ATP, ADP or AMP) after 5 h of culture of MSCs in the presence of either ectonucleotidase specific inhibitors (POM-1 or APCP) or human mAbs (anti-CD39 and anti-CD73) are shown. Asterisk indicates significant (P < 0.001) differences compared to the either CD39 or CD73 basal expression. e The expression of the CD39 and CD73 ectonucleotidases on MSCs cultured during 5 h alone with culture medium (CM) or in the presence of nucleotides (ATP, ADP and AMP), inhibitors (POM-1 and APCP) and mAbs (anti-CD39 or anti-CD73) is shown. Data are representative of three independent experiments, and the mean ± SEM are shown

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