Adipose tissue is inextricably linked to nutritional balance and metabolic diseases, and the way of adipocytes differentiation will directly affect the health of adipose tissue. Preadipocytes are cells that are present at adipose depot and restricted to becoming mature adipocytes specificity. Their favorable differentiation ability to differentiate into healthy mature adipocytes as well as undergo hyperplasia (the expansion of adipose tissue by de novo adipocytes) rather than hypertrophy (the expansion of adipose tissue by increasing the size of already being adipocytes) is crucial for regenerative medicine and obesity-related diseases.Despite much effort has focused on the factors and mechanism about preadipocyte's adipogenic differentiation, the precise regulatory mechanism is still not completely clear. Based on the related research in recent years, this review discusses the ambiguous definition, the origin and terminal differentiation of preadipocytes in brief, summarizes the surface markers of preadipocytes in detail which include stem cell surface markers (such as CD29, CD34, CD38 and SCA1), perivascular markers (such as PDGFRα and PDGFRβ), ZFP423 (zinc-finger protein 423), Pref-1/DLK1 (preadipocyte factor 1). The deep look at preadipocytes provides new ideas for clinical diagnosis and treatment. The clinical application potential of preadipocytes in the treatment of soft tissue defects, obesity-related metabolic diseases, tumors (breast cancer and prostate cancer), and wound healing is further discussed in this review.