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Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen

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Abstract

Purpose

It has been reported that cancer stem cells (CSCs) can be isolated from primitive neuroectodermal tumor (PNET) specimens. Moreover, mesenchymal stem-like cells (MSLCs) have been isolated from Korean glioma specimens. Here, we tested whether tumor spheres and MSLCs can be simultaneously isolated from a single PNET specimen, a question that has not been addressed.

Methods

We isolated single-cell suspensions from PNET specimens, then cultured these cells using methods for MSLCs or CSCs. Cultured cells were analyzed for surface markers of CSCs using immunocytochemistry and for surface markers of bone marrow-derived mesenchymal stem cells (BM-MSCs) using fluorescence-activated cell sorting (FACS). Tumor spheres were exposed to neural differentiation conditions, and MSLCs were exposed to mesenchymal differentiation conditions. Possible locations of MSLCs within PNET specimens were determined by immunofluorescence analysis of tumor sections.

Results

Cells similar to tumor spheres and MSLCs were independently isolated from one of two PNET specimens. Spheroid cells, termed PNET spheres, were positive for CD133 and nestin, and negative for musashi and podoplanin. PNET spheres were capable of differentiation into immature neural cells and astrocytes, but not oligodendrocytes or mature neural cells. FACS analysis revealed that adherent cells isolated from the same PNET specimen, termed PNET-MSLCs, had surface markers similar to BM-MSCs. These cells were capable of mesenchymal differentiation. Immunofluorescence labeling indicated that some CD105+ cells might be closely related to endothelial cells and pericytes.

Conclusion

We showed that both tumor spheres and MSLCs can be isolated from the same PNET specimen. PNET-MSLCs occupied a niche in the vicinity of the vasculature and could be a source of stroma for PNETs.

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Acknowledgments

This research was supported by grants from the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010-0004506), and the National R&D Program for Cancer Control, Ministry for Health, Welfare and Family Affairs, Republic of Korea (1020340).

Conflict of interest

The authors declare no potential conflicts of interest.

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Authors and Affiliations

  1. Department of Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea

    Jiyong Kwak

  2. Department of Neurosurgery, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea

    Hye-Jin Shin & Yong-Kil Hong

  3. Department of Pathology, Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea

    Se-Hoon Kim

  4. Department of Neurosurgery, Severance Hospital, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul, 120-752, Republic of Korea

    Jin-Kyoung Shim, Ji-Hyun Lee, Eui-Hyun Kim, Eun-Kyung Park, Jong Hee Chang, Sun Ho Kim, Dong-Seok Kim & Seok-Gu Kang

  5. Department of Radiology, Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea

    Yong-Min Huh

  6. Department of Chemistry, Hanyang University, 17 Haendang-dong, Seongdong-gu, Seoul, 133-791, Republic of Korea

    Su-Jae Lee

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  1. Jiyong Kwak
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Corresponding authors

Correspondence to Su-Jae Lee or Seok-Gu Kang.

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Jiyong Kwak and Hye-Jin Shin contributed equally to this manuscript.

Su-Jae Lee and Seok-Gu Kang contributed equally to this manuscript.

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Kwak, J., Shin, HJ., Kim, SH. et al. Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen. Childs Nerv Syst 29, 2229–2239 (2013). https://doi.org/10.1007/s00381-013-2201-x

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  • DOI: https://doi.org/10.1007/s00381-013-2201-x

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