The male sterile lines of diploid Chinese cabbage have been widely investigated for theoretical studies and practical uses. In tetraploid Chinese cabbage， improvement of fertility and seed setting rate in selfbred lines was favorable， while study of the male sterile lines remained un-investigated. In this study， a sterile source（ CMS hou36gao） with new Brassica rapa L. cytoplasmic male sterile（ CMS） gene was selected from the diploid Chinese cabbage， and subjected for chromosome doubling using colchicine treatment. By multiple consecutive cycles with tetraploid inbred lines D571 and D574， in conjugation with cytogenetically identifying the number of chromosomes， two tetraploid lines CMS-D571 and CMS-D574 that showed cytoplasmic male sterile were produced. Both lines showed sterility rate up to 100% with high seed setting rate and genetic stability. This study raised an example by using the cytoplasmic male sterility in tetraploid Chinese cabbage breeding， and established an experimental pipeline to generate the cytoplasmic tetraploid male sterile lines. Collectively， this work laid a foundation for innovation and practical use of male sterile lines in tetraploid Chinese cabbage.