Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines

Isaac O. Karikari, Christopher L. Gilchrist, Liufang Jing, David A. Alcorta, Jun Chen, William J. Richardson, Mostafa A. Gabr, Richard D. Bell, Michael J. Kelley, Carlos A. Bagley, Lori A. Setton

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Object. Chordoma cells can generate solid-like tumors in xenograft models that express some molecular characteristics of the parent tumor, including positivity for brachyury and cytokeratins. However, there is a dearth of molecular markers that relate to chordoma tumor growth, as well as the cell lines needed to advance treatment. The objective in this study was to isolate a novel primary chordoma cell source and analyze the characteristics of tumor growth in a mouse xenograft model for comparison with the established U-CH1 and U-CH2b cell lines. Methods. Primary cells from a sacral chordoma, called "DVC-4, " were cultured alongside U-CH1 and U-CH2b cells for more than 20 passages and characterized for expression of CD24 and brachyury. While brachyury is believed essential for driving tumor formation, CD24 is associated with healthy nucleus pulposus cells. Each cell type was subcutaneously implanted in NOD/SCID/IL2Rγnull mice. The percentage of solid tumors formed, time to maximum tumor size, and immunostaining scores for CD24 and brachyury (intensity scores of 0-3, heterogeneity scores of 0-1) were reported and evaluated to test differences across groups. Results. The DVC-4 cells retained chordoma-like morphology in culture and exhibited CD24 and brachyury expression profiles in vitro that were similar to those for U-CH1 and U-CH2b. Both U-CH1 and DVC-4 cells grew tumors at rates that were faster than those for U-CH2b cells. Gross tumor developed at nearly every site (95%) injected with U-CH1 and at most sites (75%) injected with DVC-4. In contrast, U-CH2b cells produced grossly visible tumors in less than 50% of injected sites. Brachyury staining was similar among tumors derived from all 3 cell types and was intensely positive (scores of 2-3) in a majority of tissue sections. In contrast, differences in the pattern and intensity of staining for CD24 were noted among the 3 types of cell-derived tumors (p < 0.05, chi-square test), with evidence of intense and uniform staining in a majority of U-CH1 tumor sections (score of 3) and more than half of the DVC-4 tumor sections (scores of 2-3). In contrast, a majority of sections from U-CH2b cells stained modestly for CD24 (scores of 1-2) with a predominantly heterogeneous staining pattern. Conclusions. This is the first report on xenografts generated from U-CH2b cells in which a low tumorigenicity was discovered despite evidence of chordoma-like characteristics in vitro. For tumors derived from a primary chordoma cell and U-CH1 cell line, similarly intense staining for CD24 was observed, which may correspond to their similar potential to grow tumors. In contrast, U-CH2b tumors stained less intensely for CD24. These results emphasize that many markers, including CD24, may be useful in distinguishing among chordoma cell types and their tumorigenicity in vivo.

Original languageEnglish (US)
Pages (from-to)386-393
Number of pages8
JournalJournal of Neurosurgery: Spine
Volume21
Issue number3
DOIs
StatePublished - Jan 1 2014

Fingerprint

Chordoma
Heterografts
Cell Line
Neoplasms
Staining and Labeling
SCID Mice

Keywords

  • Brachyury
  • CD24
  • Chordoma
  • Oncology
  • U-CH1
  • U-CH2b
  • Xenograft

ASJC Scopus subject areas

  • Surgery
  • Neurology
  • Clinical Neurology

Cite this

Karikari, I. O., Gilchrist, C. L., Jing, L., Alcorta, D. A., Chen, J., Richardson, W. J., ... Setton, L. A. (2014). Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines. Journal of Neurosurgery: Spine, 21(3), 386-393. https://doi.org/10.3171/2014.4.SPINE13262

Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines. / Karikari, Isaac O.; Gilchrist, Christopher L.; Jing, Liufang; Alcorta, David A.; Chen, Jun; Richardson, William J.; Gabr, Mostafa A.; Bell, Richard D.; Kelley, Michael J.; Bagley, Carlos A.; Setton, Lori A.

In: Journal of Neurosurgery: Spine, Vol. 21, No. 3, 01.01.2014, p. 386-393.

Research output: Contribution to journalArticle

Karikari, IO, Gilchrist, CL, Jing, L, Alcorta, DA, Chen, J, Richardson, WJ, Gabr, MA, Bell, RD, Kelley, MJ, Bagley, CA & Setton, LA 2014, 'Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines', Journal of Neurosurgery: Spine, vol. 21, no. 3, pp. 386-393. https://doi.org/10.3171/2014.4.SPINE13262
Karikari, Isaac O. ; Gilchrist, Christopher L. ; Jing, Liufang ; Alcorta, David A. ; Chen, Jun ; Richardson, William J. ; Gabr, Mostafa A. ; Bell, Richard D. ; Kelley, Michael J. ; Bagley, Carlos A. ; Setton, Lori A. / Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines. In: Journal of Neurosurgery: Spine. 2014 ; Vol. 21, No. 3. pp. 386-393.
@article{19e902dfc4f04b0a8c2b89273b1f70ef,
title = "Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines",
abstract = "Object. Chordoma cells can generate solid-like tumors in xenograft models that express some molecular characteristics of the parent tumor, including positivity for brachyury and cytokeratins. However, there is a dearth of molecular markers that relate to chordoma tumor growth, as well as the cell lines needed to advance treatment. The objective in this study was to isolate a novel primary chordoma cell source and analyze the characteristics of tumor growth in a mouse xenograft model for comparison with the established U-CH1 and U-CH2b cell lines. Methods. Primary cells from a sacral chordoma, called {"}DVC-4, {"} were cultured alongside U-CH1 and U-CH2b cells for more than 20 passages and characterized for expression of CD24 and brachyury. While brachyury is believed essential for driving tumor formation, CD24 is associated with healthy nucleus pulposus cells. Each cell type was subcutaneously implanted in NOD/SCID/IL2Rγnull mice. The percentage of solid tumors formed, time to maximum tumor size, and immunostaining scores for CD24 and brachyury (intensity scores of 0-3, heterogeneity scores of 0-1) were reported and evaluated to test differences across groups. Results. The DVC-4 cells retained chordoma-like morphology in culture and exhibited CD24 and brachyury expression profiles in vitro that were similar to those for U-CH1 and U-CH2b. Both U-CH1 and DVC-4 cells grew tumors at rates that were faster than those for U-CH2b cells. Gross tumor developed at nearly every site (95{\%}) injected with U-CH1 and at most sites (75{\%}) injected with DVC-4. In contrast, U-CH2b cells produced grossly visible tumors in less than 50{\%} of injected sites. Brachyury staining was similar among tumors derived from all 3 cell types and was intensely positive (scores of 2-3) in a majority of tissue sections. In contrast, differences in the pattern and intensity of staining for CD24 were noted among the 3 types of cell-derived tumors (p < 0.05, chi-square test), with evidence of intense and uniform staining in a majority of U-CH1 tumor sections (score of 3) and more than half of the DVC-4 tumor sections (scores of 2-3). In contrast, a majority of sections from U-CH2b cells stained modestly for CD24 (scores of 1-2) with a predominantly heterogeneous staining pattern. Conclusions. This is the first report on xenografts generated from U-CH2b cells in which a low tumorigenicity was discovered despite evidence of chordoma-like characteristics in vitro. For tumors derived from a primary chordoma cell and U-CH1 cell line, similarly intense staining for CD24 was observed, which may correspond to their similar potential to grow tumors. In contrast, U-CH2b tumors stained less intensely for CD24. These results emphasize that many markers, including CD24, may be useful in distinguishing among chordoma cell types and their tumorigenicity in vivo.",
keywords = "Brachyury, CD24, Chordoma, Oncology, U-CH1, U-CH2b, Xenograft",
author = "Karikari, {Isaac O.} and Gilchrist, {Christopher L.} and Liufang Jing and Alcorta, {David A.} and Jun Chen and Richardson, {William J.} and Gabr, {Mostafa A.} and Bell, {Richard D.} and Kelley, {Michael J.} and Bagley, {Carlos A.} and Setton, {Lori A.}",
year = "2014",
month = "1",
day = "1",
doi = "10.3171/2014.4.SPINE13262",
language = "English (US)",
volume = "21",
pages = "386--393",
journal = "Journal of Neurosurgery: Spine",
issn = "1547-5654",
publisher = "American Association of Neurological Surgeons",
number = "3",

}

TY - JOUR

T1 - Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines

AU - Karikari, Isaac O.

AU - Gilchrist, Christopher L.

AU - Jing, Liufang

AU - Alcorta, David A.

AU - Chen, Jun

AU - Richardson, William J.

AU - Gabr, Mostafa A.

AU - Bell, Richard D.

AU - Kelley, Michael J.

AU - Bagley, Carlos A.

AU - Setton, Lori A.

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Object. Chordoma cells can generate solid-like tumors in xenograft models that express some molecular characteristics of the parent tumor, including positivity for brachyury and cytokeratins. However, there is a dearth of molecular markers that relate to chordoma tumor growth, as well as the cell lines needed to advance treatment. The objective in this study was to isolate a novel primary chordoma cell source and analyze the characteristics of tumor growth in a mouse xenograft model for comparison with the established U-CH1 and U-CH2b cell lines. Methods. Primary cells from a sacral chordoma, called "DVC-4, " were cultured alongside U-CH1 and U-CH2b cells for more than 20 passages and characterized for expression of CD24 and brachyury. While brachyury is believed essential for driving tumor formation, CD24 is associated with healthy nucleus pulposus cells. Each cell type was subcutaneously implanted in NOD/SCID/IL2Rγnull mice. The percentage of solid tumors formed, time to maximum tumor size, and immunostaining scores for CD24 and brachyury (intensity scores of 0-3, heterogeneity scores of 0-1) were reported and evaluated to test differences across groups. Results. The DVC-4 cells retained chordoma-like morphology in culture and exhibited CD24 and brachyury expression profiles in vitro that were similar to those for U-CH1 and U-CH2b. Both U-CH1 and DVC-4 cells grew tumors at rates that were faster than those for U-CH2b cells. Gross tumor developed at nearly every site (95%) injected with U-CH1 and at most sites (75%) injected with DVC-4. In contrast, U-CH2b cells produced grossly visible tumors in less than 50% of injected sites. Brachyury staining was similar among tumors derived from all 3 cell types and was intensely positive (scores of 2-3) in a majority of tissue sections. In contrast, differences in the pattern and intensity of staining for CD24 were noted among the 3 types of cell-derived tumors (p < 0.05, chi-square test), with evidence of intense and uniform staining in a majority of U-CH1 tumor sections (score of 3) and more than half of the DVC-4 tumor sections (scores of 2-3). In contrast, a majority of sections from U-CH2b cells stained modestly for CD24 (scores of 1-2) with a predominantly heterogeneous staining pattern. Conclusions. This is the first report on xenografts generated from U-CH2b cells in which a low tumorigenicity was discovered despite evidence of chordoma-like characteristics in vitro. For tumors derived from a primary chordoma cell and U-CH1 cell line, similarly intense staining for CD24 was observed, which may correspond to their similar potential to grow tumors. In contrast, U-CH2b tumors stained less intensely for CD24. These results emphasize that many markers, including CD24, may be useful in distinguishing among chordoma cell types and their tumorigenicity in vivo.

AB - Object. Chordoma cells can generate solid-like tumors in xenograft models that express some molecular characteristics of the parent tumor, including positivity for brachyury and cytokeratins. However, there is a dearth of molecular markers that relate to chordoma tumor growth, as well as the cell lines needed to advance treatment. The objective in this study was to isolate a novel primary chordoma cell source and analyze the characteristics of tumor growth in a mouse xenograft model for comparison with the established U-CH1 and U-CH2b cell lines. Methods. Primary cells from a sacral chordoma, called "DVC-4, " were cultured alongside U-CH1 and U-CH2b cells for more than 20 passages and characterized for expression of CD24 and brachyury. While brachyury is believed essential for driving tumor formation, CD24 is associated with healthy nucleus pulposus cells. Each cell type was subcutaneously implanted in NOD/SCID/IL2Rγnull mice. The percentage of solid tumors formed, time to maximum tumor size, and immunostaining scores for CD24 and brachyury (intensity scores of 0-3, heterogeneity scores of 0-1) were reported and evaluated to test differences across groups. Results. The DVC-4 cells retained chordoma-like morphology in culture and exhibited CD24 and brachyury expression profiles in vitro that were similar to those for U-CH1 and U-CH2b. Both U-CH1 and DVC-4 cells grew tumors at rates that were faster than those for U-CH2b cells. Gross tumor developed at nearly every site (95%) injected with U-CH1 and at most sites (75%) injected with DVC-4. In contrast, U-CH2b cells produced grossly visible tumors in less than 50% of injected sites. Brachyury staining was similar among tumors derived from all 3 cell types and was intensely positive (scores of 2-3) in a majority of tissue sections. In contrast, differences in the pattern and intensity of staining for CD24 were noted among the 3 types of cell-derived tumors (p < 0.05, chi-square test), with evidence of intense and uniform staining in a majority of U-CH1 tumor sections (score of 3) and more than half of the DVC-4 tumor sections (scores of 2-3). In contrast, a majority of sections from U-CH2b cells stained modestly for CD24 (scores of 1-2) with a predominantly heterogeneous staining pattern. Conclusions. This is the first report on xenografts generated from U-CH2b cells in which a low tumorigenicity was discovered despite evidence of chordoma-like characteristics in vitro. For tumors derived from a primary chordoma cell and U-CH1 cell line, similarly intense staining for CD24 was observed, which may correspond to their similar potential to grow tumors. In contrast, U-CH2b tumors stained less intensely for CD24. These results emphasize that many markers, including CD24, may be useful in distinguishing among chordoma cell types and their tumorigenicity in vivo.

KW - Brachyury

KW - CD24

KW - Chordoma

KW - Oncology

KW - U-CH1

KW - U-CH2b

KW - Xenograft

UR - http://www.scopus.com/inward/record.url?scp=84908129704&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84908129704&partnerID=8YFLogxK

U2 - 10.3171/2014.4.SPINE13262

DO - 10.3171/2014.4.SPINE13262

M3 - Article

C2 - 24905390

AN - SCOPUS:84908129704

VL - 21

SP - 386

EP - 393

JO - Journal of Neurosurgery: Spine

JF - Journal of Neurosurgery: Spine

SN - 1547-5654

IS - 3

ER -