Lactate Influences the Gene Expression Profile of Human Mesenchymal Stem Cells (hMSC) in a Dose Dependant Manner

Carl-Christoph Schneider, Atesch Ateschrang, Ingmar Königsrainer, Jörg Glatzle, Sarah Bühler, Richard Schaefer, Hinnak Northoff, Alfred Königsrainer, Derek Zieker
2012 Cellular Physiology and Biochemistry  
Background/Aims: Wounds, especially non-healing wounds are characterized by elevated tissue lactate concentrations. Lactate is known for being able to stimulate collagen synthesis and vessel growth. Lately it has been shown that lactate, in vivo, plays an important role in homing of stem cells. With this work we aimed to show the influence of lactate on the gene expressionprofile of human mesenchymal stem cells (hMSC). Materials and Methods: hMSCs were obtained from bone marrow and
more » ... with fluorescence-activated cell sorting (FACS) analysis. Subsequently the hMSCs were treated with either 0, 5, 10 and 15 mM lactate (pH 7,4) for 24 hours. RNA Isolation from stimulated hMSCs and controls was performed. The Microarray analysis was performed using AffymetrixHuGene 1.0 ST Gene Chip. Selected targets were subsequently analysed using quantitative real time PCR (RTq-PCR). Results: We were able to show that lactate in moderate concentrations of 5 respectively 10 mM leads to an anti-inflammatory, anti-apoptotic but growth and proliferation promoting gene expression after 24 h. In contrast, high lactate concentrations of 15 mM leads to the opposed effect, namely promoting inflammation and apoptosis. Hypoxia induced genes did not show any significant regulation. Contrary to expectation, we were not able to show any significant regulation of candidates associated with glycolysis. Conclusion: We were able to show that lactate alters gene expression but does not change the cell phenotype, which might be helpful for further investigations of new treatment strategies for chronic non-healing wounds as well as tumor-therapy and neuronal plasticity. Pre-incubation for 300 s at 95°C for 1 cycle (20°C/s), amplification for 42 cycles each with denaturation for 10 s at 95°C (20°C/s), annealing for 5 s at 60°C (20°C/s) and extension for 15 s at 72°C (20°C/s), melting curve 15 s at 50°C (20°C/s) then continuous ascent with 0,1°C/s until 95°C for 1 cycle, followed by cooling 30 s at 40°C for 1 cycle (20°C/s). Analysis was carried out using the relative quantification [10] and the relative expression software tool (REST©) [11] . Human hydroxymethyl-bilane synthase (HMBS) was used as reference gene for the relative quantification [10]. 1555 Schneider/Ateschrang/Königsrainer et al.: Lactate and hMSC Cellular Physiology and Biochemistry Cellular Physiology and Biochemistry isoemzyme M2 (PKM2), even though lactate is the metabolic end-product of this pathway [45]. Only a moderate up-regulation of phosphoglycerat kinase 1 (PGK1) and PKM2at 15 mM lactate could be found. Conclusion We are aware of the fact that these are gene expression rates and that the levels of the corresponding proteins need to be further investigated since various regulatory processes may occur during translation and post-translational modification. Nevertheless our data suggest that lactate is able to modulate the gene expression of hMSCs involved in wound healing in a concentration dependent manner. Furthermore lactate might be involved in the regulation of processes like tumorgenesis and neuronal plasticity and growth. These data could be helpful for investigating new treatment strategies for chronic non-healing wounds as well as tumor therapy and neuronal plasticity or regeneration.
doi:10.1159/000343342 pmid:23234875 fatcat:zfi5cpjlr5cwtooiklsxujbdre