Influence of tumor microenvironment on culture and identification of breast cancer stem cell-like microspheres

Yufen Zhu, Meijing Ren, Feng Gu, Li Fu

Research output: Contribution to journalArticle

Abstract

Objective: To investigate the influence and significance of tumor microenvironment in breast cancer stem-cell culture and identification. Methods: Cells isolated from primary breast cancer tissues were cultured in vitro in a serum-free medium PCM-2 and in the supernatant of cultured fibroblasts. The MCF-7 breast cancer cell line was used as the control group. The status of the microspheres was observed, and the proliferative capacity of the cells was detected by methyl thiazolyl tetrazolium assay. The expression of stem cell and epithelial-mesenchymal markers were detected by real-time reverse transcription polymerase chain reaction. Results: The diameter of microspheres in PCM-2 gradually increased with prolonged incubation time (t=4.996, P=0.002). The cells in the supernatant of cultured fibroblasts increased daily and mostly exhibited a spindle cell growth. The growth rate of primary breast cancer cells was faster in the supernatant of cultured fibroblasts than in PCM-2 (P=0.004). Compared with the case of cells in the supernatant of cultured fibroblasts, aldehyde dehydrogenase 1 was upregulated in the primary breast cancer cells cultured in serum-free medium PCM-2, whereas E-cadherin and Vimentin were downregulated. Conclusion: Serum-free culture can be one of the best methods for enriching breast cancer stem cell-like mammospheres. The tumor micro-environment serves a vital function in the growth and development of tumor cells and in the evolution of breast cancer.

Original languageEnglish (US)
Pages (from-to)1417-1421
Number of pages5
JournalChinese Journal of Clinical Oncology
Volume41
Issue number22
DOIs
StatePublished - Nov 30 2014

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Tumor Microenvironment
Neoplastic Stem Cells
Microspheres
Breast Neoplasms
Fibroblasts
Serum-Free Culture Media
Vimentin
Cadherins
Growth
Mesenchymal Stromal Cells
Growth and Development
Reverse Transcription
Cultured Cells
Neoplasms
Down-Regulation
Cell Culture Techniques
Cell Line
Polymerase Chain Reaction
Control Groups
Serum

Keywords

  • Breast cancer
  • Breast cancer stem cells
  • Mammospheres
  • Serum-free culture
  • Tumor microenvironment

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Influence of tumor microenvironment on culture and identification of breast cancer stem cell-like microspheres. / Zhu, Yufen; Ren, Meijing; Gu, Feng; Fu, Li.

In: Chinese Journal of Clinical Oncology, Vol. 41, No. 22, 30.11.2014, p. 1417-1421.

Research output: Contribution to journalArticle

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abstract = "Objective: To investigate the influence and significance of tumor microenvironment in breast cancer stem-cell culture and identification. Methods: Cells isolated from primary breast cancer tissues were cultured in vitro in a serum-free medium PCM-2 and in the supernatant of cultured fibroblasts. The MCF-7 breast cancer cell line was used as the control group. The status of the microspheres was observed, and the proliferative capacity of the cells was detected by methyl thiazolyl tetrazolium assay. The expression of stem cell and epithelial-mesenchymal markers were detected by real-time reverse transcription polymerase chain reaction. Results: The diameter of microspheres in PCM-2 gradually increased with prolonged incubation time (t=4.996, P=0.002). The cells in the supernatant of cultured fibroblasts increased daily and mostly exhibited a spindle cell growth. The growth rate of primary breast cancer cells was faster in the supernatant of cultured fibroblasts than in PCM-2 (P=0.004). Compared with the case of cells in the supernatant of cultured fibroblasts, aldehyde dehydrogenase 1 was upregulated in the primary breast cancer cells cultured in serum-free medium PCM-2, whereas E-cadherin and Vimentin were downregulated. Conclusion: Serum-free culture can be one of the best methods for enriching breast cancer stem cell-like mammospheres. The tumor micro-environment serves a vital function in the growth and development of tumor cells and in the evolution of breast cancer.",
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