ABSTRACT
The use of relevant, accessible, and easily reproducible preclinical models of diffuse gliomas is a prerequisite for the development of successful therapeutic approaches to their treatment. Here we studied the gene expression profile of 3D spheroids in a comparison with 2D cell cultures and tissue strains of diffuse high-grade gliomas. Using real time PCR, we evaluated the expression of Gfap, Cd44, Pten, S100b, Vegfa, Hif1a, Sox2, Melk, Gdnf, and Mgmt genes playing an important role in the progression of gliomas and regulating tumor cell proliferation, adhesion, invasion, plasticity, apoptosis, DNA repair, and recruitment of tumor-associated cells. Gene expression analysis showed that 3D spheroids are more similar to tumor tissue strains by the expression levels of Gfap, Cd44, and Pten, while the expression levels of Hif1a and Sox2 in 3D spheroids did not differ from those of 2D cell cultures, the expression levels S100b and Vegfa in 3D spheroids was higher than in other models, and the expression levels of Melk, Gdnf, and Mgmt genes changed diversely. Thus, 3D spheroid model more closely mimics the tumor tissue than 2D cell culture, but still is not the most relevant, probably due to too small size of spheroids, which does not allow reproducing hypoxia and apoptotic and necrotic processes in the tumor tissue.
ABSTRACT
The study was carried out using a novel rat model developed in our laboratory, namely16 mm diameter circular excisional wounds were generated on the abdomen which resulted in minimal scarring. Restoration of the skin integrity was completed by day 60 after the wounding surgery. By this time, regenerates on the abdomen were stronger than on the back (at, respectively, 58 and 17.4 % of the tensile strength of the intact skin at corresponding location) and the ratio of type I and type III collagens in regenerates on the abdomen reached the level of intact skin at the same location. On days 3 to 14, the ratio of Mmp9/Timp1 expression levels on the abdomen was higher than on the back. On days 20 and 30, the Mmp9/Timp1 ratio on the abdomen was identical to the level of intact skin, whereas the increased MMPs expression levels on the back were maintained until day 30. It has been shown for the first time that according to functional and molecular characteristics, wound healing on the abdomen of an adult rat is more similar to complete regeneration than scarring repair of the back skin.
Subject(s)
Cicatrix/genetics , Gene Expression Regulation , Regeneration/physiology , Skin/metabolism , Surgical Wound/genetics , Wound Healing/physiology , Abdomen , Animals , Back , Cicatrix/metabolism , Cicatrix/physiopathology , Collagen Type I/genetics , Collagen Type I/metabolism , Collagen Type III/genetics , Collagen Type III/metabolism , Extracellular Matrix/chemistry , Female , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Organ Specificity , Rats , Rats, Wistar , Skin/injuries , Surgical Wound/metabolism , Surgical Wound/physiopathology , Time Factors , Tissue Inhibitor of Metalloproteinase-1/genetics , Tissue Inhibitor of Metalloproteinase-1/metabolism , Tissue Inhibitor of Metalloproteinase-2/genetics , Tissue Inhibitor of Metalloproteinase-2/metabolismABSTRACT
Differences in the gene expression profiles in resident macrophages (in particular, Kupffer cells) and monocytes were revealed. However, these differences in gene expression profiles do not allow considering resident liver macrophages as purely M2 macrophages and monocytes as purely M1 macrophages. At the same time, a significant number of the genes upregulated in Kupffer cells are associated with normal regulation of liver functions (Arg 1, Flt, iNOs, and Kng). In monocytes, the expression of genes Alox15, Alox12, Tlr2, Tlr4, Tlr7, and Tlr8 (typical functional genes of macrophages) was also upregulated in comparison with Kupffer cells.
Subject(s)
Cell Lineage/genetics , Kupffer Cells/metabolism , Liver/metabolism , Monocytes/metabolism , Animals , Arachidonate 12-Lipoxygenase/genetics , Arachidonate 12-Lipoxygenase/metabolism , Arachidonate 15-Lipoxygenase/genetics , Arachidonate 15-Lipoxygenase/metabolism , Arginase/genetics , Arginase/metabolism , Biomarkers/metabolism , Gene Expression , Immunophenotyping , Kupffer Cells/classification , Kupffer Cells/cytology , Liver/cytology , Male , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Mice , Mice, Inbred BALB C , Monocytes/classification , Monocytes/cytology , Nitric Oxide Synthase Type II/genetics , Nitric Oxide Synthase Type II/metabolism , Toll-Like Receptor 2/genetics , Toll-Like Receptor 2/metabolism , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolism , Toll-Like Receptor 7/genetics , Toll-Like Receptor 7/metabolism , Toll-Like Receptor 8/genetics , Toll-Like Receptor 8/metabolism , Vascular Endothelial Growth Factor Receptor-1/genetics , Vascular Endothelial Growth Factor Receptor-1/metabolismABSTRACT
Expression of il1b, il6, il10, tnfa, hgf, tgfb, vegf, and fgf2 genes in the lungs and kidneys was examined on rat model of liver regeneration after subtotal hepatectomy. Enhanced expression of il6, il10, tnfa, hgf, and fgf2 genes was detected at the early terms after 80% liver resection.
Subject(s)
Kidney/metabolism , Lung/metabolism , Animals , Hepatectomy , Hepatocyte Growth Factor/metabolism , Interleukin-10/metabolism , Interleukin-6/metabolism , Liver Regeneration/physiology , Rats , Rats, Sprague-Dawley , Transforming Growth Factor beta/metabolism , Tumor Necrosis Factor-alpha/metabolism , Vascular Endothelial Growth Factor A/metabolismABSTRACT
A reproducible model of fetal liver regeneration was created. Resection of 20% liver was carried out in rat fetuses on day 17 of prenatal development. The organ weight was restored after 2 days at the expense of an increase in hepatocyte mitotic activity; cell hypertrophy was minor. After recovery, the cell composition of the operated liver did not differ from the control, i.e. the regeneration was organotypical.
Subject(s)
Fetus/physiology , Hepatectomy , Hepatocytes/physiology , Liver Regeneration/physiology , Models, Biological , Animals , Cell Division/physiology , Cell Proliferation , Fetus/surgery , Hepatocytes/cytology , Rats , Statistics, NonparametricABSTRACT
We studied the effect of non-selective intracoronary transplantation of bone marrow mononuclears on day 30 after acute coronary infarction on angiogenesis in rats. On days 14 and 30 after transplantation of mononuclear cells, stable formation of new vessels was observed. The number of venules considerably increased after transplantation of mononuclear cells, which was seen from increased volume density of blood vessels and their caliber. Stable vascularization after transplantation of mononuclear cells improves blood supply, which is essential for reparation of the myocardium.
Subject(s)
Bone Marrow Transplantation , Coronary Vessels/physiopathology , Myocardial Infarction/therapy , Neovascularization, Physiologic , Animals , Bone Marrow Cells/physiology , Cicatrix/physiopathology , Granulation Tissue/blood supply , Male , Myocardial Infarction/pathology , Myocardial Infarction/physiopathology , Myocardium/pathology , Rats , Rats, Sprague-Dawley , Regeneration , Venules/physiopathologyABSTRACT
We studied homing and differentiation fate of transplanted bone marrow mononuclears after non-selective intracoronary injection on day 30 after acute myocardial infarction in rats. Mononuclear cells migrated to the cicatrix zone where they differentiated into fibroblasts and myofibroblasts. Mononuclear cells did not differentiate into cardiomyocytes, endothelial cells, or smooth muscle cells of vascular media. Stimulation of angiogenesis and reparation of the myocardium was observed under these conditions.
Subject(s)
Bone Marrow Cells/physiology , Bone Marrow Transplantation , Cell Differentiation , Myocardial Infarction/therapy , Animals , Cell Tracking , Cicatrix/pathology , Coronary Vessels/physiopathology , Fibroblasts/pathology , Heart/physiology , Male , Myocardial Infarction/pathology , Myofibroblasts/pathology , Neovascularization, Physiologic , Rats , Rats, Sprague-Dawley , RegenerationABSTRACT
We studied hepatocyte proliferation in the regenerating liver of 17-day fetuses of outbred albino rats. The animals were sacrificed every 3 h over 2 days after resection of 20% liver. The number of mitoses beyond the zone of injury increased sharply 12 and 24 h after resection. The hepatocyte mitotic index in the perifocal zone did not surpass the mitotic index in regions distant from the focus of injury during any of the studied periods. No circadian rhythm of hepatocyte mitotic activity was detected for resected or intact fetal liver. The injury caused virtually no changes in hepatocyte mitosis phase ratio in the operated compared to intact liver, which attested to stable course of mitosis. The weight of fetal liver recovered at the expense of enhanced mitotic activity of hepatocytes in the entire liver.
Subject(s)
Cell Proliferation , Hepatocytes/physiology , Liver Regeneration/physiology , Animals , Circadian Rhythm , Fetus/metabolism , Fetus/physiology , Hepatectomy , Hepatocytes/metabolism , Liver/embryology , Mitosis , Mitotic Index , RatsABSTRACT
Heart injury was inflicted to outbred albino rat fetuses on day 16 of prenatal development and cardiomyocyte proliferation was evaluated in the left ventricular zones adjacent to the wound and distant from it. The mitotic index of experimental animals was higher than in intact controls only for zones distant from the injury on day 10 after the intervention. The percentage of pathological metaphases gradually increased in all experimental groups, which causes doubt in the viability of postmitotic cardiomyocytes.
