C-Kit expression as a feature of functional differentiation of progenitor cells

The resulted findings and the analysis of C-kit expression during human prenatal development allow referring the receptor of stem cells factor to the markers of committed precursor cells, the expression of which coincides with the functional differentiation of precursor cells. In the human pancreas there is a common C-kit-positive progenitor cell of and#945;- and and#946;-cells of the islets of Langerhans, which remains in the organ after birth

Influence of long-term cultivation and cryopreservation on phenotype of rat hepatic stellate cells

Many organs and tissues have been reported to harbor regional stem cells. One of the candidates for this role in the liver is hepatic stellate cells (HSC). Cultivation of cells in vitro may lead to changes in their morphology and phenotype. The risk of changes increases with long-term cultivation (passage 5 and above), as well as with cryopreservation of cells. Freezing allows storage of isolated cell; however, the method can influence cells phenotype and properties. The aim of research was to study phenotype of rat HSC during long-term cultivation and cryopreservation. Three cultures of HSC (I - after cryopreservation, II - long-term culture, passage 6, and III - control, passage 4) were stained immunocytochemically with antibodies to desmin, -SMA, CK19, Ki-67; quantitative real-time PCR analysis was also performed to evaluate desmin and -SMA gene expression. It was observed that morphology of cells was similar in all three groups. In groups with long-term cultivation and cryopreserA134-A13

Histochemical analysis of self-organizing 3D spheroid-like cultures of adipose-derived mesenchymal stem cells

Histochemical staining demonstrated fusion of cells with formation of syncytium-like structures. On the periphery cells were polygonal, round, had epithelial morphology. Groups of cells organized in line inside spheroid remained elongated AD-MSC shape. Most of the cells did not proliferate. There were no migrating SMA+ cells. Polygonal cells on the periphery and a few cells inside the spheroid were desmin+. Interestingly, peripheral and elongated shape cells expressed epithelial marker CK-19+. Probably, within spheroid there was spontaneous mesenchymal-epithelial transdifferentiation. When spheroids were placed in a new culture dish we observed explantational outgrowth of spindle-shaped AD-MSC suggesting that cells within spheroids were alive, able to outgrow and revert through epithelial-mesenchymal transdifferentiation back to MSC phenotype. Investigation was funded by RFBR grant 18-04-01133 and supported by Program of Competitive Growth of KFU

Postnatal development of the microstructure of cortical GABAergic synapses and perineuronal nets requires sensory input

The brain synaptic circuitry is formed as a result of pre-defined genetic programs and sensory experience during postnatal development. Perineuronal nets ensheath synaptic boutons and control several crucial features of the synapse physiology. Formation of the perineuronal net microstructure during the brain development remains largely unstudied. Here we provide a detailed quantitative description of the 3-dimensional geometry of the synapse and the surrounding perineuronal net in the mouse somatosensory cortex layer IV. We compare the morphology of the synapse+perineuronal net complex in the adult brain formed under normal conditions or in the whisker shaving model of somatosensory deprivation. We demonstrate that the sensory deprivation causes flattening of the 3D PNN mesh geometry and reduction of the VGAT-positive cluster volume in presynaptic boutons. These results reveal a mechanism of the sensory input-dependent synapse morphogenesis during the brain development.Peer reviewe

Parenchymal and nonparenchymal cellular proliferation dynamics after partial hepatectomy with and without 2-Acetylaminofluorene injection in rats

Restoration of liver after partial hepatectomy (PH) depends on proliferation dynamics of parenchymal and nonparenchymal liver cell populations. To study separately proliferation impact of nonparenchymal cells it is recommended to inhibit hepatocytes proliferation by injection of acetoaminofluorene (AAF). The aim of the research was to study dynamics of parenchymal and nonparenchymal cells proliferation after PH w/wo AAF injection. Experimental groups were 1) PH; 2) PH with intraperitoneal AAF injection (0,2ml of 1.5g/100ml). Liver paraffin slices (1,5,7,14,21,28 days after transplantation) were stained with antibodies against proliferation marker Ki-67. AAF injection inhibits proliferation of parenchymal and nonparenchymal cells, has severe effect on central vein region, leads to reduction of liver regeneration (cellular dystrophy and degeneration on 28th days). Work supported by Program of Competitive Growth of KFU.A13

Artificial microvesicles: new perspective on healing tendon wounds

Tendons have a limited capacity to repair both naturally and following clinical interventions. Damaged tissue often presents with structural and functional differences, adversely affecting animal performance, mobility, health and welfare. Advances in cell therapies have started to overcome some of these issues, however complications such as the formation of ectopic bone remain a complication of this technique. Regenerative medicine is therefore looking towards future therapies such as the introduction of microvesicles (MVs) derived from stem cells (SCs). The aim of the present study was to assess the characteristics of artificially derived MVs, from equine mesenchymal stem cells (MSCs), when delivered to rat tendon cells in vitro and damaged tendons in vivo. The initial stages of extracting MVs from equine MSCs and identifying and characterising the cultured tendon stem/progenitor cells (TSCs) from rat Achilles tendons were undertaken successfully. The horse MSCs, and the rat tendon cells, were both capable of differentiating in three directions: adipogenic, osteogenic and chondrogenic pathways. The artificially derived equine MVs successfully fused with the TSC membranes, and no cytotoxic or cytostimulating effects were observed. In addition, co-cultivation of TSCs with MVs lead to stimulation of cell proliferation and migration, and cytokine VEGF and Fractalkine expression levels were significantly increased. These experiments are the first to show that artificially derived MVs exhibited regeneration-stimulating effects in vitro, and that fusion of cytoplasmic membranes from diploid cell lines originating from different species was possible. Explorations in vivo showed accelerated regeneration of injury tendons after introduction of the MVs into damaged areas. The results from the studies performed indicated obvious positive modifying effects following the administration of MVs. This represents the initial successful steps required prior to translating this regenerative medicine technique into clinical trials, such as for tendon repair in injured horses

Cytokine Storm Combined with Humoral Immune Response Defect in Fatal Hemorrhagic Fever with Renal Syndrome Case, Tatarstan, Russia

Hemorrhagic fever with renal syndrome (HFRS) is endemic in Tatarstan, where thousands of cases are registered annually. Puumala orthohantavirus is commonly detected in human case samples as well as in captured bank voles, the rodent hosts. The pathogenesis of HFRS is still not well described, although the cytokine storm hypothesis is largely accepted. In this study, we present a comprehensive analysis of a fatal HFRS case compared with twenty four non-fatal cases where activation of the humoral and cellular immune responses, pro-inflammatory cytokines and disturbed blood coagulation were detected using immunological, histological, genetic and clinical approaches. Multiple organ failure combined with disseminated intravascular coagulation syndrome and acute renal failure was the cause of death. Decreased Interleukin (IL)-7 and increased IL-18, chemokine (C-C motif) ligand (CCL)-5, stem cell growth factor (SCGF)-b and tumor necrosis factor-beta (TNF-β) serum levels were found, supporting the cytokine storm hypothesis of hantavirus pathogenesis