OBJECTIVETo establish subseries cell lines from single, cancer cell of Tca8113M1 cell line and detect the cancer stem cell markers in the different subseries cell lines.

METHODSThe subseries cell lines from single cancer cell of Tca8113M1 cell line were established by limiting dilution assay in vitro. The characteristic of tumorigenicity and CD44, CD184, extracellular soluble antigen (ESA) of the cancer stem cell markers were detected by xenotransplantation and flow cytometry respectively.

RESULTSTotal 192 single cells of Tca8113M1 cell line were cultured and were deposited as one cell per well. There were 12 subpopulations origin from 192 single cells spheroid cultures. The ratio was 6.25% (12/192). In the different subpopulations, the tumorigenicity and expression of CD44 and ESA were at high levels, but the expression of CD184 was in different level. There were three kinds morphology of colonies derived from single cancer cells, holoclone, meroclone and paraclone. Cell line could be derived from carcinoma cell holoclones by cell culture. Meroclone and paraclone did not exist in cell culture in vitro.

CONCLUSIONTongue cancer stem cell may exist in Tca8113M1 cell line, cell line can be established and holoclone is the origin of cell line. This is a novel approach to the identification and enrichment for cancer stem cell.