Cited 0 times in

122 19

Ideal sphere-forming culture conditions to maintain pluripotency in a hepatocellular carcinoma cell lines

Authors
 Seon Ok Min ; Sang Woo Lee ; Kyung Sik Kim ; Seon Young Bak 
Citation
 Cancer Cell International, Vol.15 : 95, 2015 
Journal Title
 Cancer Cell International 
ISSN
 1475-2867 
Issue Date
2015
Abstract
BACKGROUND: Cancer stem cells (CSCs) constitute 1-2 % of cancer tissue and are a major cause of tumor metastasis and recurrence. The culture environment is important in maintaining CSCs in vitro. Sphere formation is one method of culturing CSCs. In this study, we identified and validated optimal culture conditions for sphere formation in hepatocellular carcinoma cells. METHODS: Huh7 and HepG2 cells were plated in three different media types and were allowed to form spheres. To confirm the pluripotency of sphere cells, the expression of stem cell markers was evaluated. EpCAM, Connexin 32, and Connexin 43 expression was evaluated using reverse transcription-polymerase chain reaction (RT-PCR). The expression of E-cadherin, β-catenin, CD90, and CD133 was evaluated using immunocytochemistry. Flow cytometry was performed to confirm the presence of stem cell markers CD133 and Connexin 32. RESULTS: Cells maintained in medium containing growth factors ((DMEM(+))GF) showed greater cell proliferation than cells in media with fetal bovine serum (FBS) (DMEM(+)FBS) or without FBS (DMEM(-)FBS). Cells cultured in DMEM(+)FBS medium exhibited greater proliferation after 20 days in culture than cells cultured under the other two conditions. Cells cultured in DMEM(-)FBS medium did not proliferate; therefore, this condition was removed from further analysis. RT-PCR and flow cytometry showed that sphere-forming cells cultured in DMEM(+)GF and DMEM(+)FBS media had similar expression of stem cell markers. CONCLUSION: Therefore, growth factor-free medium is an adaptable, efficient, and cost-effective tool for in vitro cultivation of CSCs.
URI
http://ir.ymlib.yonsei.ac.kr/handle/22282913/141709
DOI
10.1186/s12935-015-0240-y
Appears in Collections:
1. 연구논문 > 1. College of Medicine > Dept. of Surgery
Yonsei Authors
사서에게 알리기
  feedback
Files in This Item:
T201504170.pdfDownload
Export
RIS (EndNote)
XLS (Excel)
XML

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse