نیما دهدیلانی

Sheep satellite cells more than satellite cells of the rat and mouse are similar to human satellite cells. These cells are widely used in the modeling and treatment of diseases like heart insufficiency, neurological diseases, muscular dystrophy, cerebral cell transplantation for the treatment of migraines, screening, and the production of new drugs. This study was aimed to isolate and culture primary satellite cells (PSCs) obtained from sheep fetus, and perform clonal expansion of transfected PSCs. Skeletal muscle tissues of hind limbs were collected from sheep fetuses obtained from a local abattoir. After enzymatic digestion, flasks were replaced after 3 hours to isolate non-myogenic cells, such as fibroblasts. After six days, the cells were differentiated to myoblasts. Using a differentiation medium containing the horse serum, myotube cells were observed in the flask, indicating that the cultured cells were satellite cells. The mRNA expression of the PAX7 gene was used to confirm the presence of satellite cells. In addition, the results showed that satellite cells grow in a culture medium containing 5% FBS without differentiation, while 10% FBS initiates their differentiation.

The purpose of the present study was to isolate, culture expand and characterize canine bone marrow derived mesenchymal stem cells. Bone marrow aspirates of 15 adult male dogs were collected to this end and their mononuclear cells isolated by centrifugation and cultured in standard media. The adherent cells were isolated and their mesenchymal origin was confirmed at 3rd passage by cellular morphology, expression of surface antigens and differentiation to osteogenic and adipogenic lineage. After 4 days, spindle shaped fibroblast like cells which were apparently bone marrow derived mesenchymal stem cells appeared in culture medium and their numbers increased over time. The cells reached 3rd passage with over 75% confluent after a mean of 22.89±5.75 days. Flow cytometric analysis revealed that the cells negatively expressed CD34 and CD45 antigens while positively expressing CD44 and CD105 antigens. Differentiation into osteogenic and adipogenic lineage had taken place after one month culture in induction medium. VDR, COL1A1, BGLAP and SPARC gene expression indicated that mesenchymal stem cells isolated from canine bone marrow had differentiated into osteogenic lineage. These findings can form the basis of any forthcoming clinical studies involving the use of canine mesenchymal stem cells particularly in the field of bone and cartilage regeneration.

The effect of Baicalin، a PPARy activator، was investigated on erythroid differentiation of CD133+ cord blood hematopoietic stem cells. The effect of PPARγ agonists (Baicalin and Troglitazone) were evaluated in erythropoiesis. CD133+ cells were isolated from human cord blood using MACS technology and cultured in the medium containing PPARγ agonists in a dose-dependent manner with SCF (20 ng/mL) and Epo (4. 5 u/mL). The morphology of CD133+ cells، cell surface markers (Transferrin receptor and glycophorin A)، and the formation of erythroid colony were assessed with microscopicanalysis، flow cytometry، and colony forming assay، respectively. Microscopic and flow cytometric analysis revealed the differentiation of CD133+ cord blood hematopoietic stem cells to erythroid lineage. Flow cytometry showed that both of the PPARγ agonists were able to diminish significantly Transferrin receptor and glycophorin A positive cell population. The inhibitory effect of PPARγ agonists on erythroid differentiation was dose-dependent. The colonies formation of erythroid was suppressed by both of PPARy agonists; hence، the effect of troglitazone was markedly higher. The inhibitory effect of Troglitazone on colonies formation of Erythroid was also markedly higher than Bicaline. Our findings demonstrated that PPARy agonists modulates erythroid differentiation of CD133+ hematopoietic stem cell; therefore، they might play an important role in regulating normal erythropoiesis in vivo. The inhibitory effect of Baicalin on erythropoiesis should be noted because this herbal remedy mostly is used to treat a wide range of diseases.