The Xi(0) muon semileptonic decay has been observed for the first time with nine identified events using the KTeV beam line and detector at Fermilab. The decay is normalized to the Xi(0) beta decay mode and yields a value for the ratio of decay rates Gamma(Xi(0)->Sigma(+)mu(-)nu(mu))/Gamma(Xi(0)->Sigma(+)e(-)nu(e)) of [1.8(-0.5)(+0.7)(stat)+/- 0.2(syst)]x10(-2). This is in agreement with the SU(3) flavor symmetric quark model.