Abstract Pulmonary mesenchymal stem cells (PMSCs) are considered important in therapeutic and regenerative responses to lung injury. Despite extensive studies on the human, porcine, sheep, mouse and rat, it was found that PMSCs, which can been performed on goose, have not been well explored. Elucidation of cell differentiation, proliferation, which have not yet been well described, may promote the development of injured lung therapy. In our study, we firstly disclosed biological characteristics of spindle-shaped PMSCs from pulmonary tissues of goose embryos. Growth kinetics and counting kit-8 (CCK8) assay were employed for proliferative activity evaluation. When appropriately induced, PMSCs could differentiate into osteoblasts, adipocytes, chondrocytes, type II alveolar epithelial cells (ATII), which demonstrated that PMSCs have cross-embryonic layer differentiation potential. Besides, undifferentiated PMSCs consistently expressed MSCs associated phenotypes, such as CD166, CD90, CD44, CD29, CD71 and CD105, as identified by immunohistochemistry and RT-PCR. Karyotype analysis demonstrated that PMSCs possessed stable diploid karyotype. Collectively, we successfully established applicable methods for isolation, culture, identification and characterization of goose PMSCs which suggested a potential therapeutic use in regenerative therapy. (PMSCs injury human porcine sheep rat explored proliferation described therapy study spindleshaped spindle shaped embryos kit8 kit 8 kit- CCK8 CCK (CCK8 evaluation induced osteoblasts adipocytes chondrocytes ATII, ATII , (ATII) crossembryonic cross embryonic Besides phenotypes CD166 CD CD90 CD44 CD29 CD7 CD105 RTPCR. RTPCR RT PCR. PCR RT-PCR karyotype Collectively isolation culture (CCK (ATII CD16 CD9 CD4 CD2 CD10 CD1