الفهرس | Only 14 pages are availabe for public view |
Abstract Mesenchymal stem cells have been shown to exist in cord blood. Although MSCs have been described by a subset of surface antigens after expansion, little is known about the cell surface phenotype of undifferentiated MSCs. Human umbilical cord blood stem cells able to differentiate into hepatocytes and fibroplasts in vitro. The present study is conducted to obtain an HSC-enriched population (lineage-negative and positive CD34 cells) from the UCB and characterize their proliferation and differentiation capacities under some physiological conditions available and easy for application in both basic and clinical research. The present study showed human umbilical cord blood stem cell differentiation; human umbilical cord blood mononuclear cells were separated by MACs technology. Fibroblast and hepatocyte cell extract were added in the inducing groups. The expansion and differentiation of expression of hepatic and MSC markers in differentiated cells was analyzed by reverse-transcription polymerase chain reaction, and compared with control. The present study showed that stem progenitor cells purification were performed by phenotypic analysis in Flow Cytometry using CD34- FITC, human kit showed cell purity (76.12±4.02) % (Fig.5). The new selected CD34+ cells were spherical and remains in its viable state and forming colonies when growth medium were resuspended in 10% and changed every 3 days for 10 to 14 days of incubation at 37°C and 5% CO2 incubator (Fig.6) Following treatment with fibroblast cell extract, the majority of CD34+ cell morphology are spherical shape, some cells behave adherent growth, protruding and spindle cells were seen (Fig.7).The spherical shape of CD34+ cells treated with fibroblast cell extract were significantly amplified by adding CD34-, whereas the cytoplasm of the round cells was enlarged, the spindle-shaped cells had differentiated into fibroblast-like cells during the time course (Fig.8). The treatment of the newly selected CD34+ cells with CD34- and hepatocyte cell lyste , the spherical shaped cells forms regenerative embryonic bodies differentiated into polygonal cells characteristic to hepatocyte cells(Fig.9,10). All treated groups expressed optical density of RNA pattern compared to control. In conclusion, both of hepatocyte and fibroblast cell homogenate were sufficient for expansion of UCB CD34+ -cells and co-transplantation of MSC with UCB CD34+ cells, promoting engraftment of UCB CD34+cells. |