The invention provides a method for cultivating isolated microspore of cucumber, belonging to the field of biological technology. The method of the invention comprises the steps as follows: taking buds, identifying the development period of the microspore, pre-disposing, disinfecting the buds, isolating the microspore with an extrusion method by using B<5>-13 extracting solution, filtrating, centrifuging and purifying the microspore by cell screens with the size of 76mum, carrying out static dark cultivation at the temperature of 25 DEG C till the embryoid can be seen by naked eyes, forming cotyledonary embryoid by surging dark cultivation under the frequency of 60r/min<-1> subsequently, forming regenerated plants or rootless seedlings on MS-BA embryoid germination culture mediums, and finally gaining entire regenerated plants by inducing the roots on the rooting culture medium MS-IBA. The method of the invention can gain haploid or double haploid. The double haploid is good seedling raising material and can be used for the selection of new type or improvement of type, thus greatly shortening the seedling raising period and improving the selection efficiency; the double haploid is also ideal material for carrying out the molecular marking such as AFLP, RFLP, RAPD, and the like, the drawing and researching of genetic maps, the species evolution research, genetic analysis, plant gene cloning selection, etc.