biostudies-literaturePark SCNIDDK NIH HHSNCI NIH HHS1506-14https://www.ebi.ac.uk/biostudies/studies/S-EPMC4499771biostudies-literatureUnknown24(13)A large number of cancer stem cells (CSCs) have been isolated and identified; however, none has been cultured in an unlimited manner in vitro without losing tumorigenicity and multipotency. In this study, we successfully clonogenically cultured a newly identified CD34+ liver CSC (LCSC) on feeder cells up to 22 passages (to date) without losing CSC property. Cloned CD34+ LCSC formed a round packed morphology and it could also be cryopreserved and recultured. Stem cell markers, CD34, CD117, and SOX2; normal liver stem cell markers, alpha fetoprotein, CK19, CK18, and OV6; putative CSC markers, CD44, CD133, EpCAM, and CD90; as well as CD31 were expressed in cloned CD34+ LCSC. SOX2 was the major factor in maintaining this LCSC before colonization, and interestingly, OCT4, SOX2, NAONG, Klf4, c-MStem cells and developmentClonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro.PMC4499771DK075415P30 CA093373Zeng CZhao MPark SCZhang YRamsamooj RNguyen NTEun JRTschudy-Seney BZhou HZern MATheise NDDuan YfalseClonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro.A large number of cancer stem cells (CSCs) have been isolated and identified; however, none has been cultured in an unlimited manner in vitro without losing tumorigenicity and multipotency. In this study, we successfully clonogenically cultured a newly identified CD34+ liver CSC (LCSC) on feeder cells up to 22 passages (to date) without losing CSC property. Cloned CD34+ LCSC formed a round packed morphology and it could also be cryopreserved and recultured. Stem cell markers, CD34, CD117, and SOX2; normal liver stem cell markers, alpha fetoprotein, CK19, CK18, and OV6; putative CSC markers, CD44, CD133, EpCAM, and CD90; as well as CD31 were expressed in cloned CD34+ LCSC. SOX2 was the major factor in maintaining this LCSC before colonization, and interestingly, OCT4, SOX2, NAONG, Klf4, c-M2015-01-01T00:00:00Z2015 Jul2026-07-15T01:05:23.015Z2019-03-27T01:55:00ZS-EPMC44997712586758310.1089/scd.2015.0022