Effect of human adipose-derived mesenchymal stem-C on the differentiation of monocytes

The ideal stem cell for use in functional tissue engineering needs to be abundantly available, harvested with minimal morbidity, differentiated reliably down various pathways and able to be transplanted safely and efficaciously. Adult human adipose tissue contains a population of mesenchymal stem cells; adipose-derived stem cells, which seem to fulfil most, if not all, of these criteria. In this work, we investigated the immunogenicity properties of human adipose-derived mesenchymal stem cells [HAMSCs] and their effect on monocytes differentiation. The HAMSCs have been isolated and specified. Human peripheral blood mononuclear cells [PBMCs] were isolated and passed through a column with magnetic beads coated with anti-CD14 antibody. CD14+ ve cells were isolated and cultured independently or co-cultured with HAMSCs in the presence of cytokines [IL-4, Granulocyte-macrophage colony-stimulating factor [GM-CSF]] to induce their differentiation into dendritic cells [DCs]. Their further maturation was induced by LPS added on the 6[th] day of culture. The major part of the independently cultured cells [CD14+ ve] was found to express the markers which are considered to be specific for the mature dendritic cells such as Human leukocyte antigen-DR [HLA-DR] [40.44%] and low percentage of cells [6.9%]. Nevertheless dendritic cells of monocyte origin [mDCs] co-cultured with HAMSCs showed significant shifts in the pattern of surface markers. The percentage of HLA-DR cells was much lower [6.44%] compared to control cultures [p < 0.001]. Similarly, the secretion of IL-10 by DCs was up-regulated in co-cultures of HAMSCs and DCs. The results show that human adipose-tissue mesenchymal stem cells [HAMSCs] could inhibit the differentiation of the blood monocytes into dendritic cells

Role of 5-azacytidine in the differentiation of rat bone marrow mesenchymal stem cells into cardiomyogenic cells

Increasing attention is being paid to the use of mesenchymal stem cells [MSCs] for treatment of human diseases such as myocardial infarction. To study the differentiation of the bone marrow mesenchymal stem cells [BM-MSCs] into cardiomyogenic cells using 5-azacytidine. Forty adult male albino rats were used in this study. BM-MSCs were isolated and cultured in a complete Dulbecco's modified Eagle's medium containing 1% antibiotics and 10% fetal bovine serum [the control group]. Second passaged cells were treated with 10micro moI/I 5-azacytidine for 72h. Then, the medium was removed and kept in a 5-azacytidine-free medium for 4 weeks [the 5-azacytidine-treated group]. The adherent cells of both groups were examined using a phase-contrast microscope and a transmission electron microscope. Expressions of cytoskeleton protein desmin and cardiac muscle-specific cardiac troponin T were assessed by immunohistochemistry. BM-MSCs of the control group were spindled and star shaped with multiple processes and vesicular nuclei. After adding 5-azacytidine for 1 week, the cells showed multinucleation. On the second week, the cells formed stick-like structures. The cells showed extensive cytoplasmic striations in the third week. Finally, in the fourth week, the cells formed myotube-like structures. Immunohistochemical staining of cells of the 5-azacytidine-treated group revealed a positive immune reaction for desmin and cardiac troponin-T. Ultrastructural examination of the 5-azacytidine-treated group revealed that the cells were elongated with central oval large nuclei. The mitochondria were elongated with well developed cristae. There were abundant free ribosomes and extensive dilated rough endoplasmic reticulum. Myofibrils started to appear in the peripheral part of the cytoplasm and T-tubules appeared. MSCs can be differentiated in vitro by 5-azacytidine into cardiomyogenic cells, which are important for repairing infracted myocardium

Comparison of behavior, morphology and morphometry of equine and canine adipose derived mesenchymal stem cells in culture / Comparación del comportamiento, morfología y morfometría del tejido adiposo equino y canino derivado de células madre mesenquimales en cultivo

Recent studies revealed multipotent properties of fat tissue isolated mesenchymal stem cells. These cells are successfully used as therapeutic factor for many locomotive disorders, being even more effective than stem cells from bone marrow. Isolated and cultured, AD-MSCs were observed, photographed and measured to compare cells from two different species.

Estudios recientes han revelado propiedades pluripotentes del tejido graso aislado de células madre mesenquimales. Estas células se utilizan con éxito como factor terapéutico para muchos trastornos locomotores, siendo aún más eficaz que las células madre de médula ósea. Aisladas y cultivadas, AD-MSC se observaron, fotografiaron y midieron comparar células de dos especies diferentes.

Bio treatment protects rat marrow-derived mesenchymal stem cell culture against the TNF-a induced apoptosis

This study is an attempt to examine the anti apoptotic effects of BIO on rat MSC culture. Rat marrow primary cell culture was established and exposure groups were defined; cultures with 0.01, 0.1, 1 micro M BIO. Cells cultured without BIO treatment were used as controls. During culture expantion, the average doubling time, as an index of the rate of cell growth, were determined and compared. To examine whether or not BIO is able to protect MSCs against apoptosis, the passaged-3 cells from each group were induced to undergo apoptosis with the addition of TNF-alpha [Tumor necrotic factor-alpha]. Three days after, the cultures were quantified in terms of the percentages of apoptotic cells using either the Tunnel or Annexin V staining method. Marrow cells cultivated with 0.1 and 1 micro M BIO appeared to expand at a significantly more rapid rate than the 0.01 micro M BIO and the control cultures [p < 0.05]. Tunnel staining indicated that in 1 micro M BIO-treated groups, there were lower percentages of apoptotic nuclei than in groups with other concentrations of BIO [p < 0.05]. The BIO protective effect appeared to be dose-dependent in that the cultures with high BIO content possessed less apoptotic nuclei. The results obtained by Annexin staining were in agreement with the results of Tunnel staining. The Annexin method additionally takes into account the early apoptotic cells which are not detectable by the Tunnel method. Taken together, it seems that cultivation with BIO could both increase the growth rate of marrow cells and protect MSCs against induced apoptosis