Transdifferentiation Of The Sca-1 Positive Cells From The Murine Fetal Liver And Application

Object: To explore the potential and behavior of transdifferentiation of Sca-l+ cells from murine fetal liver as well as their repairing capacity in acute renal failure. Method: The single cell suspension was prepared of murine fetal liver of pregnancy of 14.5day; cell sex was determined by quick PCR and male cells were collected, and Sca-l+ cells of them were separated with MACS technology. 2x104 cells of male Sca-l+ cells were transplanted into female syngeneic mouse irradiated with lethally dose of Y ray by tail vein. During 2-6 months, the kidneys, livers, lungs, stomach and small intestinal tissue were taken out and paraffin slices of them were made. The Y-chromosome cells were traced by fluorescence in situ hybridization (FISH), at the same time; the tissue characteristics of them were detected by immunochemistry. In order to detect the repairing capacity of Sca-l+ cells from fetal liver, the model of acute renal failure was made with glycerol. The urea nitrogen and creatine in serum were monitored after 10 H g/kg of G-CSF was injected into model mice. Furthermore, In order to reduce the interference of female hematopoietic stem cells in vivo, female bone marrow would be changed into male one with elimination of female bone marrow by radiation and transplantation of male Sca-l+ cells from fetal liver, which needed time of 8 weeks. The acute renal failure model of female mice with this male bone marrow was made again, and 72 hours later, Sca-l+ cells from fetal liver were transplanted into them. 8 weeks later, the mice were killed and kidneys were took out to be made tissue slices for detecting of FISH and immunochemistry, the transdifferentiation frequency of Sca-l+ cells from fetal liver and bone marrow were calculated and compared. Results: The male cells were found on sections of kidneys, liver, lung, gastric and smallintestinal tissue of female mice irradiated with lethally Y ray and transplanted with male sca-l+ cells, frequency of differentiation were (4.5+0.5)%, (0.9+0.1)%, (1.9+0.6)%, (6.1 0.5)% and (7.61 2.3)% respectively, phenotype characteristics of male renal tissue was RCA+/CYP1A2+/Vimentin+/CD457F4/80". There was a big increase of frequency of male cells from (4.5+₀.5)% to (8.58 1.34)% on sections of female mouse of acute renal failure compared with single dose of radiation, and there appeared a double male cell arrangement on area of renal tubules on section. There was much bigger increase of frequency of male on section of female mouse (18.13+₁.91)%, bone marrow sex of which had become male, of acute renal failure, and there appeared ring arrangement of several male cells on area of renal tubule on sections. The transdifferentiation frequency of Sca-l+ cells from fetal liver was higher than bone marrow on model of acute renal failure, as follows (3.55+0.51)% and (1.35 0.09)% respect.Conclusion: The Sca-l+ cells from fetal liver have the capacity differentiated into kidneys, liver, and lung, gastric and small intestinal tissue. The higher frequency of transdifferntiation would be obtained in acute renal failure. The transdifferentiation potential of Sca-l+ cells from fetal liver was much bigger than bone marrow.