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Abstract Consented umbilical cord blood cells from male full-term deliveries were obtained. Mononuclear layer from the human umbilical cord blood cells were isolated by fractionation on Ficoll solution. Cells were then incubated in conical tubes at 37°C in a 5% CO2 atmosphere for 7 days in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with FBS, penicillin, streptomycin, amphotrecin and hepatocyte growth factor. The viability of cultured cell was determined by using trypan blue staining and the cultured cells were impregnated with ferumoxides injectable solution (feridex). Then, immunohistochemical staining was done to the cultured cells for human AFP to detect the hepatocytes progenitor cells. For the in vivo study, 120 adult female albino rats, weighting 200-250 gm were used in this study. The rats were divided into three groups: Control group (n=20) , Liver injury group without stem cells transplantation (n=50): Received, D-galactosamine hydrochloride intraperitoneal injection (300mg/kg) and Liver injury with stem cell transplantation group (n=50). AST, ALT, Albumin and bilirubin were estimated before and after liver injury in rats. In vitro study: Using HGF, It was found that the transdifferentiation of the umbilical cord blood cells to hepatocyte progenitor occurred on the 7th day after culture. The number of the hepatocyte progenitor cells stained by AFP was 12.2 ± 3.42 cells /HPF. Determination of albumin in the culture supernatant was estimated on the 7th day and found to be 1.03 ± 0.16 μg /ml. In vivo study: Stem cell transplantation to the animal models was done by intrasplenic injection under light ether anesthesia . An improvement of AST, ALT, albumin and bilirubin activities were observed at one month (44.2 ± 8.87, 196.8 ± 39.87, 3.4 ± 0.38 and 0.27 ± 0.01 respectively) after intrasplenic stem cells transplantation. After the 3 months, the values of AST, ALT, albumin and bilirubin were found to be near the control values (33.6 ± 4.27, 151.4 ± 35.94, 3.94 ± 0.11 and 0.24 ± 0.02 respectively). After one and three months of UCB stem cells transplantation into the rats with liver injury, both spleen and liver showed AFP positive cells. The mean number of AFP positive cells in the spleen was (14.8 ± 7.14 cells/HPF). The area % of AFP was used to denote the quantity of AFP expression within the hepatocyte progenitor. The area % of AFP at 1 month was (3.773 ± 1.154), while it was (0.95 ± 0.404) at 3 months. Histological examination showed strong positive PAS stained cells in the spleen at the 3rd month after hepatocytes progenitor cells transplantation. Ferumoxides impregnated progenitor cells could be detected after intrasplenic transplantation by prussian blue stain in the spleen. At the 3rd month, the number of prussian blue stained cells in the spleen (37.9 ± 9.04 cells /HPF) was approximately near or equal to the number of AFP positive cells. |