Increasing evidence suggests that many types of cancers contain a population of cells that display stem cell properties. These cells are called cancer stem cells (CSCs),which are closely related to tumor initiation,growth,metastasis and chemoresistance. CSCs are also found in esophageal squamous cell carcinoma (ESCC). These cells are characterized by potential of self-renewal and differentiation,tumor formation in nude mice and chemotherapy resistance,and thus may play an important role in targeted cancer therapies. Current methods for culturing and sorting CSCs in ESCC mainly include fluorescence activated cell sorting (FACS),magnetic activated cell sorting (MACS),suspension culture,and side population (SP) cell sorting. In this review,we focus on current research methods for CSCs in ESCC,their biological characteristics and areas for improvement. We believe that a combination of multiple cell-surface makers is needed for research of CSCs in ESCC.