Investigation On The Main Factors Affecting Prk?c? Knockout Mice Establishment In Blood System

?Objective?We try to establish Prk?c? knock-out mice in the blood system by fetal liver cell transplantation and investigate the main factors affecting model establishment.?Methods?The fetal liver isolated from the pregnant mouse embryos were fully grinded for cell suspension and one-to-one transplanted into lethally-irradiated wild type C57BL/6 recipient mice.The tails of fetuses were taken for Prk?c? genotyping.The expression level of Prk?c? protein in blood system was detected by western blot.The influence of irradiation(8.5Gy,9.5Gy,10.5 Gy),embryonic age(E13.5,E15.5,E17.5,E19.5)of the donors,the way of transfusion(in peri-orbital sinus,tail vein injection)on Prk?c? knockout mice in the blood system has been compared.Survival was monitored for 30 days after transplantation.Kaplan-Meier method was used to display survival curve.?Results?The percent survival(in order 100%,91%,78%)of the mice irradiated(8.5Gy,9.5Gy,10.5Gy)and transplanted with the fetal liver cell has the statistically significant difference(P<0.05).The mice irradiated(8.5Gy)and transplanted with Prk?c?-/-FLCs has Prk?c? protein in blood system,the mice irradiated(9.5Gy,10.5Gy)and transplanted with Prk?c?-/-FLCs has no Prk?c? protein.The influence of irradiation(8.5Gy)compared with irradiation(9.5Gy,10.5Gy)has statistically significant difference on the gene homogeneity(P < 0.05).The percent survival of the mice transplanted with the fetal liver cell from embryonic age(E13.5,E15.5,E17.5,E19.5)are(in order 89%,91%,88%,89%),they have no statistically significant difference with each other(P>0.05).The percent survival of the mice with the way of transfusion(peri-orbital sinus,tail vein injection,respectively)is 91%,80%,they have statistically significant difference(P<0.05).?Conclusions?Using the following conditions that irradiation(9.5Gy),fetal liver cell from the embryonic age(E13.5,E15.5,E17.5,E19.5),the way of transfusion in peri-orbital sinus can establish the blood system of Prk?c? knockout model simply and effectively.