Construction of megakaryocyte/platelet Creg1 knockout mice and induced differentiation of megakaryocytes from fetal liver
Objective Megakaryocyte/platelet specific Creg1(cellular repressor of E1A-stimulated genes 1)knockout mice were con-structed,and induce megakaryocyte differentiation in mouse fetal liver.Methods Female Creg1flox/flox mice were bred with male plate-let factor4(Pf4)-Cre+mice to obtain Creg1flox/wt PF4-Cre+mice.Further mating yielded megakaryocyte/platelet-specific Creg1 knock-out mice(Creg1flox/flox Pf4-Cre+,Creg1-/-for short).Genotypes were identified by polymerase chain reaction and agarose gel electropho-resis.Fetal liver of Creg1flox/flox and Creg1-/-mice with gestational age of about 15 days was taken,fetal liver cells were cultured in vitro for 4 days,and throbopoietin was added to observe the difference in the morphology and size of megakaryocytes between the two groups under light microscope.Results In this study,Creg1-/-mice were successfully constructed.Compared with Creg1flox/flox mice,the ex-pression of Creg1 mRNA in megakaryocytes of Creg1-/-mice was significantly decreased,with statistical significance(P<0.05).Com-pared with Creg1flox/flox mice,the diameter of megakaryocytes in fetal liver of Creg1-/-mice decreased significantly on the 4th day of in-duction differentiation,and the difference was statistically significant(P<0.05).Conclusion Creg1-/-mice,as experimental tools to study the pathogenesis of megakaryocyte and platelet-related diseases,may reveal important pathophysiological mechanisms that have not yet been discovered and clarified,and provide theoretical support for clinical treatment of platelet diseases.
Cellular repressor of E1A-stimulated genes 1MegakaryocytesBlood plateletsDifferentiation
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