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

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

Effect of curcumin and gliotoxin on rat liver myofibroblasts culture

Since 1990s, when it was demonstrated by Hammel and others, that liver fibrosis is reversible, researchers and physicians actively search for new antifibrotic therapies. In recent years knowledge of liver fibrosis pathophysiology has greatly advanced and new cellular and molecular mechanisms were described. The cells that determine extracellular matrix components distribution are myofibroblasts, but their origin is diverse. They can be activated hepatic stellate cells (HSC), portal fibroblasts (PF) or circulating mesenchymal stem cells of bone marrow. Activation of HSC and PF and their fibrogenic potential is upregulated by transcription factor NF-B. Among large number of substrates to inhibit NF-B and induce apoptosis we chose curcumin and gliotoxin. Primarily in current work we optimized the explantation culture method for isolation of hepatic myofibroblasts and received two different cultures - myofibroblasts of HSC and PF origin. Exposition of 50 ?M curcumin and 0,1 ?M gliotoxi

Influence of adenoviral transduction with Adv5-optHGF-RFP on phenotype of hepatic stellate cells

Adenoviral transduction of hepatic stellate cells is applicable for short time stimulation of genes expression, safety for phenotype and cells proliferation. At the same time adenoviral transduction with growth factors genes leads to expression of hepatoblasts markers

Effect of host liver cell proliferation on homing and phenotype of transplanted hepatic stellate cells after partial hepatectomy in rats

After PH first proliferate hepatocytes, then start nonparenchymal HSC and the last react cholangiocytes. Transplanted RFP+cells during the first week retained morphology and phenotype of hepatocytes, after 14 days they were RFP+/Desmin+ HSC, after 21 days found as RFP+/CK19+ cholangiocytes. Localization and phenotype of transplanted cells is strongly related with dynamic of host hepatic cell population proliferation

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