Objective To explore techniques of isolating embryonic stem-like cells (ESLCs) from mouse blastocyst inner cell mass and the utilization of ESLCs to generate chimerical mice. Methods\ ESLCs from mouse blastocyst inner cell masses of 3\^5 days were isolated as donor' cells to inject receptors'blastocysts to generate chimerical mice by micromanipulating techniques, and all those blastocysts were transfered to the uterine of pseudopregnant female mice to gernerate chimerical mice. Results All the ESLCs isolated from 36 inner cell masses were injected into 256 blastocysts of Kunming mice ,and those blastocysts were transferred to uterine of 32 pseudopregnant female mice, 2 chimerical mice from 12 gernerations were obtained. Conclusion \ Technique of isolating ESLCs was a new approach in generating chimerical animals, especially in some animals in which Esc lines were not constructed.