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Child's Nervous System
Published in: Child's Nervous System 12/2013

01-12-2013 | Original Paper

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

Authors: Jiyong Kwak, Hye-Jin Shin, Se-Hoon Kim, Jin-Kyoung Shim, Ji-Hyun Lee, Yong-Min 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

Published in: Child's Nervous System | Issue 12/2013

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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.
Literature
1.
Cho KT, Wang KC, Kim SK, Shin SH, Chi JG, Cho BK (2002) Pediatric brain tumors: statistics of SNUH, Korea (1959–2000). Childs Nerv Syst 18(1–2):30–37PubMedCrossRef
2.
Dirks PB, Harris L, Hoffman HJ, Humphreys RP, Drake JM, Rutka JT (1996) Supratentorial primitive neuroectodermal tumors in children. J Neurooncol 29(1):75–84PubMedCrossRef
3.
Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8(4):315–317PubMedCrossRef
4.
Dvorak HF (1986) Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing. N Engl J Med 315(26):1650–1659PubMedCrossRef
5.
Enguita-German M, Schiapparelli P, Rey JA, Castresana JS (2010) CD133+ cells from medulloblastoma and PNET cell lines are more resistant to cyclopamine inhibition of the sonic hedgehog signaling pathway than CD133− cells. Tumour Biol 31(5):381–390PubMedCrossRef
6.
7.
Galli R, Binda E, Orfanelli U, Cipelletti B, Gritti A, De Vitis S, Fiocco R, Foroni C, Dimeco F, Vescovi A (2004) Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res 64(19):7011–7021PubMedCrossRef
8.
Gan PP, Pasquier E, Kavallaris M (2007) Class III beta-tubulin mediates sensitivity to chemotherapeutic drugs in non small cell lung cancer. Cancer Res 67(19):9356–9363PubMedCrossRef
9.
Hayatsu N, Kaneko MK, Mishima K, Nishikawa R, Matsutani M, Price JE, Kato Y (2008) Podocalyxin expression in malignant astrocytic tumors. Biochem Biophys Res Commun 374(2):394–398PubMedCrossRef
10.
Hemmati HD, Nakano I, Lazareff JA, Masterman-Smith M, Geschwind DH, Bronner-Fraser M, Kornblum HI (2003) Cancerous stem cells can arise from pediatric brain tumors. Proc Natl Acad Sci U S A 100(25):15178–15183PubMedCrossRef
11.
Howell L, Mensah A, Brennan B, Makin G (2005) Detection of recurrence in childhood solid tumors. Cancer 103(6):1274–1279PubMedCrossRef
12.
Hussein D, Punjaruk W, Storer LC, Shaw L, Ottoman R, Peet A, Miller S, Bandopadhyay G, Heath R, Kumari R, Bowman KJ, Braker P, Rahman R, Jones GD, Watson S, Lowe J, Kerr ID, Grundy RG, Coyle B (2011) Pediatric brain tumor cancer stem cells: cell cycle dynamics, DNA repair, and etoposide extrusion. Neuro Oncol 13(1):70–83PubMedCrossRef
13.
Kaneko Y, Sakakibara S, Imai T, Suzuki A, Nakamura Y, Sawamoto K, Ogawa Y, Toyama Y, Miyata T, Okano H (2000) Musashi1: an evolutionally conserved marker for CNS progenitor cells including neural stem cells. Dev Neurosci 22(1–2):139–153PubMedCrossRef
14.
Kang SG, Shinojima N, Hossain A, Gumin J, Yong RL, Colman H, Marini F, Andreeff M, Lang FF (2010) Isolation and perivascular localization of mesenchymal stem cells from mouse brain. Neurosurgery 67(3):711–720PubMedCrossRef
15.
Kim SM, Kang SG, Park NR, Mok HS, Huh YM, Lee SJ, Jeun SS, Hong YK, Park CK, Lang FF (2011) Presence of glioma stroma mesenchymal stem cells in a murine orthotopic glioma model. Childs Nerv Syst 27(6):911–922PubMedCrossRef
16.
Kim YG, Jeon S, Sin GY, Shim JK, Kim BK, Shin HJ, Lee JH, Huh YM, Lee SJ, Kim EH, Park EK, Kim SH, Chang JH, Kim DS, Kim SH, Hong YK, Kang SG, Lang FF (2013) Existence of glioma stroma mesenchymal stemlike cells in Korean glioma specimens. Childs Nerv Syst 29(4):549–563PubMedCrossRef
17.
Kong BH, Park NR, Shim JK, Kim BK, Shin HJ, Lee JH, Huh YM, Lee SJ, Kim SH, Kim EH, Park EK, Chang JH, Kim DS, Kim SH, Hong YK, Kang SG, Lang FF (2013) Isolation of glioma cancer stem cells in relation to histological grades in glioma specimens. Childs Nerv Syst 29(2):217–229PubMedCrossRef
18.
Kong BH, Shin HD, Kim SH, Mok HS, Shim JK, Lee JH, Shin HJ, Huh YM, Kim EH, Park EK, Chang JH, Kim DS, Hong YK, Kim SH, Lee SJ, Kang SG (2013) Increased in vivo angiogenic effect of glioma stromal mesenchymal stem-like cells on glioma cancer stem cells from patients with glioblastoma. Int J Oncol 42(5):1754–1762PubMed
19.
Kuhl J, Muller HL, Berthold F, Kortmann RD, Deinlein F, Maass E, Graf N, Gnekow A, Scheurlen W, Gobel U, Wolff JE, Bamberg M, Kaatsch P, Kleihues P, Rating D, Sorensen N, Wiestler OD (1998) Preradiation chemotherapy of children and young adults with malignant brain tumors: results of the German pilot trial HIT'88/'89. Klin Padiatr 210(4):227–233PubMedCrossRef
20.
Lang FF, Amano T, Hata N, Gumin J, Aldape K, Colman H (2007) Bone marrow-derived mesenchymal stem cells are recruited to and alter the growth of human gliomas [abstract]. Neuro Oncol 9:596
21.
Lang FF, Gumin J, Amano T, Hata N, Heimberger F, Marini F, Andreeff M, Aldape K, Sulman E, Colman H (2008) Tumor-derived mesenchymal stem cells in human gliomas: Isolation and biological properties [abstract]. J Clin Oncol 26(15S):2001
22.
Lennon DP, Caplan AI (2006) Isolation of human marrow-derived mesenchymal stem cells. Exp Hematol 34(11):1604–1605PubMedCrossRef
23.
Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW, Kleihues P (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol 114(2):97–109PubMedCrossRef
24.
Mareschi K, Biasin E, Piacibello W, Aglietta M, Madon E, Fagioli F (2001) Isolation of human mesenchymal stem cells: bone marrow versus umbilical cord blood. Haematologica 86(10):1099–1100PubMed
25.
Merchant TE, Fouladi M (2005) Ependymoma: new therapeutic approaches including radiation and chemotherapy. J Neurooncol 75(3):287–299PubMedCrossRef
26.
Messahel B, Ashley S, Saran F, Ellison D, Ironside J, Phipps K, Cox T, Chong WK, Robinson K, Picton S, Pinkerton CR, Mallucci C, Macarthur D, Jaspan T, Michalski A, Grundy RG, Children's Cancer Leukaemia Group Brain Tumour C (2009) Relapsed intracranial ependymoma in children in the UK: patterns of relapse, survival and therapeutic outcome. Eur J Cancer 45(10):1815–1823PubMedCrossRef
27.
Mishima K, Kato Y, Kaneko MK, Nishikawa R, Hirose T, Matsutani M (2006) Increased expression of podoplanin in malignant astrocytic tumors as a novel molecular marker of malignant progression. Acta Neuropathol 111(5):483–488PubMedCrossRef
28.
Mozzetti S, Ferlini C, Concolino P, Filippetti F, Raspaglio G, Prislei S, Gallo D, Martinelli E, Ranelletti FO, Ferrandina G, Scambia G (2005) Class III beta-tubulin overexpression is a prominent mechanism of paclitaxel resistance in ovarian cancer patients. Clin Cancer Res 11(1):298–305PubMed
29.
Paget S (1889) The distribution of secondary growths in cancer of the breast. Lancet 133:571–573CrossRef
30.
Picard D, Miller S, Hawkins CE, Bouffet E, Rogers HA, Chan TS, Kim SK, Ra YS, Fangusaro J, Korshunov A, Toledano H, Nakamura H, Hayden JT, Chan J, Lafay-Cousin L, Hu P, Fan X, Muraszko KM, Pomeroy SL, Lau CC, Ng HK, Jones C, Van Meter T, Clifford SC, Eberhart C, Gajjar A, Pfister SM, Grundy RG, Huang A (2012) Markers of survival and metastatic potential in childhood CNS primitive neuro-ectodermal brain tumours: an integrative genomic analysis. Lancet Oncol 13(8):838–848PubMedCrossRef
31.
Piccirillo SG, Combi R, Cajola L, Patrizi A, Redaelli S, Bentivegna A, Baronchelli S, Maira G, Pollo B, Mangiola A, DiMeco F, Dalpra L, Vescovi AL (2009) Distinct pools of cancer stem-like cells coexist within human glioblastomas and display different tumorigenicity and independent genomic evolution. Oncogene 28(15):1807–1811PubMedCrossRef
32.
Shin GY, Shim JK, Lee JH, Shin HJ, Lee SJ, Huh YM, Kim EH, Park EK, Kim SH, Chang JH, Kim DS, Hong YK, Kim SH, Kang SG, Lang FF (2013) Changes in the biological characteristics of glioma cancer stem cells after serial in vivo subtransplantation. Childs Nerv Syst 29(1):55–64PubMedCrossRef
33.
Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J, Dirks PB (2003) Identification of a cancer stem cell in human brain tumors. Cancer Res 63(18):5821–5828PubMed
34.
Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, Henkelman RM, Cusimano MD, Dirks PB (2004) Identification of human brain tumour initiating cells. Nature 432(7015):396–401PubMedCrossRef
35.
Timmermann B, Kortmann RD, Kuhl J, Meisner C, Dieckmann K, Pietsch T, Bamberg M (2002) Role of radiotherapy in the treatment of supratentorial primitive neuroectodermal tumors in childhood: results of the prospective German brain tumor trials HIT 88/89 and 91. J Clin Oncol 20(3):842–849PubMedCrossRef
36.
Tommasi S, Mangia A, Lacalamita R, Bellizzi A, Fedele V, Chiriatti A, Thomssen C, Kendzierski N, Latorre A, Lorusso V, Schittulli F, Zito F, Kavallaris M, Paradiso A (2007) Cytoskeleton and paclitaxel sensitivity in breast cancer: the role of beta-tubulins. Int J Cancer 120(10):2078–2085PubMedCrossRef
37.
Turner CD, Rey-Casserly C, Liptak CC, Chordas C (2009) Late effects of therapy for pediatric brain tumor survivors. J Child Neurol 24(11):1455–1463PubMedCrossRef
38.
Zhang QB, Ji XY, Huang Q, Dong J, Zhu YD, Lan Q (2006) Differentiation profile of brain tumor stem cells: a comparative study with neural stem cells. Cell Res 16(12):909–915PubMedCrossRef
Metadata
Title
Isolation of tumor spheres and mesenchymal stem-like cells from a single primitive neuroectodermal tumor specimen
Authors
Jiyong Kwak
Hye-Jin Shin
Se-Hoon Kim
Jin-Kyoung Shim
Ji-Hyun Lee
Yong-Min 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
Publication date
01-12-2013
Publisher
Springer Berlin Heidelberg
Published in
Child's Nervous System / Issue 12/2013
Print ISSN: 0256-7040
Electronic ISSN: 1433-0350
DOI
https://doi.org/10.1007/s00381-013-2201-x

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