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

Jiyong Kwak, Hye Jin Shin, SeHoon Kim, Jin Kyoung Shim, Ji Hyun Lee, yongmin Huh, Eui Hyun Kim, Eun Kyung Park, Jong Hee Chang, Sun Ho Kim, Yong Kil Hong, Dong Seok Kim, Su Jae Lee, Seok-Gu Kang

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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.

Original languageEnglish
Pages (from-to)2229-2239
Number of pages11
JournalChild's Nervous System
Volume29
Issue number12
DOIs
Publication statusPublished - 2013 Dec 1

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Primitive Neuroectodermal Tumors
Mesenchymal Stromal Cells
Neoplasms
Neoplastic Stem Cells
Fluorescent Antibody Technique
Cultured Cells
Flow Cytometry
Bone Marrow
Nestin
Pericytes
Oligodendroglia
Glioma
Astrocytes
Suspensions

All Science Journal Classification (ASJC) codes

  • Pediatrics, Perinatology, and Child Health
  • Clinical Neurology

Cite this

Kwak, Jiyong ; Shin, Hye Jin ; Kim, SeHoon ; Shim, Jin Kyoung ; Lee, Ji Hyun ; Huh, yongmin ; Kim, Eui Hyun ; Park, Eun Kyung ; Chang, Jong Hee ; Kim, Sun Ho ; Hong, Yong Kil ; Kim, Dong Seok ; Lee, Su Jae ; Kang, Seok-Gu. / Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen. In: Child's Nervous System. 2013 ; Vol. 29, No. 12. pp. 2229-2239.
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title = "Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen",
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.",
author = "Jiyong Kwak and Shin, {Hye Jin} and SeHoon Kim and Shim, {Jin Kyoung} and Lee, {Ji Hyun} and yongmin Huh and Kim, {Eui Hyun} and Park, {Eun Kyung} and Chang, {Jong Hee} and Kim, {Sun Ho} and Hong, {Yong Kil} and Kim, {Dong Seok} and Lee, {Su Jae} and Seok-Gu Kang",
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Kwak, J, Shin, HJ, Kim, S, Shim, JK, Lee, JH, Huh, Y, Kim, EH, Park, EK, Chang, JH, Kim, SH, Hong, YK, Kim, DS, Lee, SJ & Kang, S-G 2013, 'Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen', Child's Nervous System, vol. 29, no. 12, pp. 2229-2239. https://doi.org/10.1007/s00381-013-2201-x

Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen. / Kwak, Jiyong; Shin, Hye Jin; Kim, SeHoon; Shim, Jin Kyoung; Lee, Ji Hyun; Huh, yongmin; Kim, Eui Hyun; Park, Eun Kyung; Chang, Jong Hee; Kim, Sun Ho; Hong, Yong Kil; Kim, Dong Seok; Lee, Su Jae; Kang, Seok-Gu.

In: Child's Nervous System, Vol. 29, No. 12, 01.12.2013, p. 2229-2239.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen

AU - Kwak, Jiyong

AU - Shin, Hye Jin

AU - Kim, SeHoon

AU - Shim, Jin Kyoung

AU - Lee, Ji Hyun

AU - Huh, yongmin

AU - Kim, Eui Hyun

AU - Park, Eun Kyung

AU - Chang, Jong Hee

AU - Kim, Sun Ho

AU - Hong, Yong Kil

AU - Kim, Dong Seok

AU - Lee, Su Jae

AU - Kang, Seok-Gu

PY - 2013/12/1

Y1 - 2013/12/1

N2 - 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.

AB - 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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