Clonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro

Su Cheol Park, Changjun Zeng, Benjamin Tschudy-Seney, Ngoc Tue Nguyen, Jong Ryeol Eun, Yanling Zhang, Rajendra Ramsamooj, Yanghong Zhang, Min Zhao, Neil D. Theise, Huaijun Zhou, Mark A Zern, YuYou Duan

Research output: Contribution to journalArticle

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Abstract

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-Myc, and Lin28 were upregulated in association with symmetric self-renewal for colony growth of CD34 + LCSC on feeder cells. Gene expression patterns of in vitro differentiation were consistent with our in vivo finding; furthermore, the tumorigenicity of cloned CD34 + LCSC was not different from uncloned CD34 + LCSC sorted from parental PLC. These results show that our cloned CD34 + LCSC maintained CSC property, including self-renewal, bipotency, and tumorigenicity after long-term culture, demonstrating that this LCSC can be cultured in an unlimited manner in vitro. Thus, establishing pure population of CSCs isolated from the patients will provide an opportunity to explore the mechanisms of tumorigenesis and cancer development, and to identify unique biomarkers presenting potential indicators of drug efficacy against CSCs for establishment of a novel strategy for cancer therapy.

Original languageEnglish (US)
Pages (from-to)1506-1514
Number of pages9
JournalStem Cells and Development
Volume24
Issue number13
DOIs
StatePublished - Jul 1 2015

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Neoplastic Stem Cells
Liver Neoplasms
Liver
Feeder Cells
Stem Cells
In Vitro Techniques
alpha-Fetoproteins
Neoplasms
Carcinogenesis
Biomarkers
Gene Expression

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Hematology

Cite this

Park, S. C., Zeng, C., Tschudy-Seney, B., Nguyen, N. T., Eun, J. R., Zhang, Y., ... Duan, Y. (2015). Clonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro. Stem Cells and Development, 24(13), 1506-1514. https://doi.org/10.1089/scd.2015.0022

Clonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro. / Park, Su Cheol; Zeng, Changjun; Tschudy-Seney, Benjamin; Nguyen, Ngoc Tue; Eun, Jong Ryeol; Zhang, Yanling; Ramsamooj, Rajendra; Zhang, Yanghong; Zhao, Min; Theise, Neil D.; Zhou, Huaijun; Zern, Mark A; Duan, YuYou.

In: Stem Cells and Development, Vol. 24, No. 13, 01.07.2015, p. 1506-1514.

Research output: Contribution to journalArticle

Park, SC, Zeng, C, Tschudy-Seney, B, Nguyen, NT, Eun, JR, Zhang, Y, Ramsamooj, R, Zhang, Y, Zhao, M, Theise, ND, Zhou, H, Zern, MA & Duan, Y 2015, 'Clonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro', Stem Cells and Development, vol. 24, no. 13, pp. 1506-1514. https://doi.org/10.1089/scd.2015.0022
Park SC, Zeng C, Tschudy-Seney B, Nguyen NT, Eun JR, Zhang Y et al. Clonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro. Stem Cells and Development. 2015 Jul 1;24(13):1506-1514. https://doi.org/10.1089/scd.2015.0022
Park, Su Cheol ; Zeng, Changjun ; Tschudy-Seney, Benjamin ; Nguyen, Ngoc Tue ; Eun, Jong Ryeol ; Zhang, Yanling ; Ramsamooj, Rajendra ; Zhang, Yanghong ; Zhao, Min ; Theise, Neil D. ; Zhou, Huaijun ; Zern, Mark A ; Duan, YuYou. / Clonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro. In: Stem Cells and Development. 2015 ; Vol. 24, No. 13. pp. 1506-1514.
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AU - Eun, Jong Ryeol

AU - Zhang, Yanling

AU - Ramsamooj, Rajendra

AU - Zhang, Yanghong

AU - Zhao, Min

AU - Theise, Neil D.

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AU - Duan, YuYou

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AB - 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-Myc, and Lin28 were upregulated in association with symmetric self-renewal for colony growth of CD34 + LCSC on feeder cells. Gene expression patterns of in vitro differentiation were consistent with our in vivo finding; furthermore, the tumorigenicity of cloned CD34 + LCSC was not different from uncloned CD34 + LCSC sorted from parental PLC. These results show that our cloned CD34 + LCSC maintained CSC property, including self-renewal, bipotency, and tumorigenicity after long-term culture, demonstrating that this LCSC can be cultured in an unlimited manner in vitro. Thus, establishing pure population of CSCs isolated from the patients will provide an opportunity to explore the mechanisms of tumorigenesis and cancer development, and to identify unique biomarkers presenting potential indicators of drug efficacy against CSCs for establishment of a novel strategy for cancer therapy.

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