The requirement for freshly isolated human colorectal cancer (CRC) cells in isolating CRC stem cells

F. Fan, Seth Bellister, J. Lu, X. Ye, D. R. Boulbes, F. Tozzi, E. Sceusi, S. Kopetz, F. Tian, L. Xia, Y. Zhou, R. Bhattacharya, L. M. Ellis

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background:Isolation of colorectal cancer (CRC) cell populations enriched for cancer stem cells (CSCs) may facilitate target identification. There is no consensus regarding the best methods for isolating CRC stem cells (CRC-SCs). We determined the suitability of various cellular models and various stem cell markers for the isolation of CRC-SCs.Methods:Established human CRC cell lines, established CRC cell lines passaged through mice, patient-derived xenograft (PDX)-derived cells, early passage/newly established cell lines, and cells directly from clinical specimens were studied. Cells were FAC-sorted for the CRC-SC markers CD44, CD133, and aldehyde dehydrogenase (ALDH). Sphere formation and in vivo tumorigenicity studies were used to validate CRC-SC enrichment.Results:None of the markers studied in established cell lines, grown either in vitro or in vivo, consistently enriched for CRC-SCs. In the three other cellular models, CD44 and CD133 did not reliably enrich for stemness. In contrast, freshly isolated PDX-derived cells or early passage/newly established CRC cell lines with high ALDH activity formed spheres in vitro and enhanced tumorigenicity in vivo, whereas cells with low ALDH activity did not.Conclusions:PDX-derived cells, early passages/newly established CRC cell lines and cells from clinical specimen with high ALDH activity can be used to identify CRC-SC-enriched populations. Established CRC cell lines should not be used to isolate CSCs.

Original languageEnglish (US)
Pages (from-to)539-546
Number of pages8
JournalBritish Journal of Cancer
Volume112
Issue number3
DOIs
StatePublished - Feb 3 2015
Externally publishedYes

Fingerprint

Neoplastic Stem Cells
Colorectal Neoplasms
Aldehyde Dehydrogenase
Cell Line
Heterografts
Cell Separation
Population
Stem Cells

Keywords

  • cancer stem cells
  • colorectal cancer
  • tumorigenicity

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

The requirement for freshly isolated human colorectal cancer (CRC) cells in isolating CRC stem cells. / Fan, F.; Bellister, Seth; Lu, J.; Ye, X.; Boulbes, D. R.; Tozzi, F.; Sceusi, E.; Kopetz, S.; Tian, F.; Xia, L.; Zhou, Y.; Bhattacharya, R.; Ellis, L. M.

In: British Journal of Cancer, Vol. 112, No. 3, 03.02.2015, p. 539-546.

Research output: Contribution to journalArticle

Fan, F, Bellister, S, Lu, J, Ye, X, Boulbes, DR, Tozzi, F, Sceusi, E, Kopetz, S, Tian, F, Xia, L, Zhou, Y, Bhattacharya, R & Ellis, LM 2015, 'The requirement for freshly isolated human colorectal cancer (CRC) cells in isolating CRC stem cells', British Journal of Cancer, vol. 112, no. 3, pp. 539-546. https://doi.org/10.1038/bjc.2014.620
Fan, F. ; Bellister, Seth ; Lu, J. ; Ye, X. ; Boulbes, D. R. ; Tozzi, F. ; Sceusi, E. ; Kopetz, S. ; Tian, F. ; Xia, L. ; Zhou, Y. ; Bhattacharya, R. ; Ellis, L. M. / The requirement for freshly isolated human colorectal cancer (CRC) cells in isolating CRC stem cells. In: British Journal of Cancer. 2015 ; Vol. 112, No. 3. pp. 539-546.
@article{e2005e903ddb4f5daf11e030ddee52c3,
title = "The requirement for freshly isolated human colorectal cancer (CRC) cells in isolating CRC stem cells",
abstract = "Background:Isolation of colorectal cancer (CRC) cell populations enriched for cancer stem cells (CSCs) may facilitate target identification. There is no consensus regarding the best methods for isolating CRC stem cells (CRC-SCs). We determined the suitability of various cellular models and various stem cell markers for the isolation of CRC-SCs.Methods:Established human CRC cell lines, established CRC cell lines passaged through mice, patient-derived xenograft (PDX)-derived cells, early passage/newly established cell lines, and cells directly from clinical specimens were studied. Cells were FAC-sorted for the CRC-SC markers CD44, CD133, and aldehyde dehydrogenase (ALDH). Sphere formation and in vivo tumorigenicity studies were used to validate CRC-SC enrichment.Results:None of the markers studied in established cell lines, grown either in vitro or in vivo, consistently enriched for CRC-SCs. In the three other cellular models, CD44 and CD133 did not reliably enrich for stemness. In contrast, freshly isolated PDX-derived cells or early passage/newly established CRC cell lines with high ALDH activity formed spheres in vitro and enhanced tumorigenicity in vivo, whereas cells with low ALDH activity did not.Conclusions:PDX-derived cells, early passages/newly established CRC cell lines and cells from clinical specimen with high ALDH activity can be used to identify CRC-SC-enriched populations. Established CRC cell lines should not be used to isolate CSCs.",
keywords = "cancer stem cells, colorectal cancer, tumorigenicity",
author = "F. Fan and Seth Bellister and J. Lu and X. Ye and Boulbes, {D. R.} and F. Tozzi and E. Sceusi and S. Kopetz and F. Tian and L. Xia and Y. Zhou and R. Bhattacharya and Ellis, {L. M.}",
year = "2015",
month = "2",
day = "3",
doi = "10.1038/bjc.2014.620",
language = "English (US)",
volume = "112",
pages = "539--546",
journal = "British Journal of Cancer",
issn = "0007-0920",
publisher = "Nature Publishing Group",
number = "3",

}

TY - JOUR

T1 - The requirement for freshly isolated human colorectal cancer (CRC) cells in isolating CRC stem cells

AU - Fan, F.

AU - Bellister, Seth

AU - Lu, J.

AU - Ye, X.

AU - Boulbes, D. R.

AU - Tozzi, F.

AU - Sceusi, E.

AU - Kopetz, S.

AU - Tian, F.

AU - Xia, L.

AU - Zhou, Y.

AU - Bhattacharya, R.

AU - Ellis, L. M.

PY - 2015/2/3

Y1 - 2015/2/3

N2 - Background:Isolation of colorectal cancer (CRC) cell populations enriched for cancer stem cells (CSCs) may facilitate target identification. There is no consensus regarding the best methods for isolating CRC stem cells (CRC-SCs). We determined the suitability of various cellular models and various stem cell markers for the isolation of CRC-SCs.Methods:Established human CRC cell lines, established CRC cell lines passaged through mice, patient-derived xenograft (PDX)-derived cells, early passage/newly established cell lines, and cells directly from clinical specimens were studied. Cells were FAC-sorted for the CRC-SC markers CD44, CD133, and aldehyde dehydrogenase (ALDH). Sphere formation and in vivo tumorigenicity studies were used to validate CRC-SC enrichment.Results:None of the markers studied in established cell lines, grown either in vitro or in vivo, consistently enriched for CRC-SCs. In the three other cellular models, CD44 and CD133 did not reliably enrich for stemness. In contrast, freshly isolated PDX-derived cells or early passage/newly established CRC cell lines with high ALDH activity formed spheres in vitro and enhanced tumorigenicity in vivo, whereas cells with low ALDH activity did not.Conclusions:PDX-derived cells, early passages/newly established CRC cell lines and cells from clinical specimen with high ALDH activity can be used to identify CRC-SC-enriched populations. Established CRC cell lines should not be used to isolate CSCs.

AB - Background:Isolation of colorectal cancer (CRC) cell populations enriched for cancer stem cells (CSCs) may facilitate target identification. There is no consensus regarding the best methods for isolating CRC stem cells (CRC-SCs). We determined the suitability of various cellular models and various stem cell markers for the isolation of CRC-SCs.Methods:Established human CRC cell lines, established CRC cell lines passaged through mice, patient-derived xenograft (PDX)-derived cells, early passage/newly established cell lines, and cells directly from clinical specimens were studied. Cells were FAC-sorted for the CRC-SC markers CD44, CD133, and aldehyde dehydrogenase (ALDH). Sphere formation and in vivo tumorigenicity studies were used to validate CRC-SC enrichment.Results:None of the markers studied in established cell lines, grown either in vitro or in vivo, consistently enriched for CRC-SCs. In the three other cellular models, CD44 and CD133 did not reliably enrich for stemness. In contrast, freshly isolated PDX-derived cells or early passage/newly established CRC cell lines with high ALDH activity formed spheres in vitro and enhanced tumorigenicity in vivo, whereas cells with low ALDH activity did not.Conclusions:PDX-derived cells, early passages/newly established CRC cell lines and cells from clinical specimen with high ALDH activity can be used to identify CRC-SC-enriched populations. Established CRC cell lines should not be used to isolate CSCs.

KW - cancer stem cells

KW - colorectal cancer

KW - tumorigenicity

UR - http://www.scopus.com/inward/record.url?scp=84922241491&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84922241491&partnerID=8YFLogxK

U2 - 10.1038/bjc.2014.620

DO - 10.1038/bjc.2014.620

M3 - Article

C2 - 25535733

AN - SCOPUS:84922241491

VL - 112

SP - 539

EP - 546

JO - British Journal of Cancer

JF - British Journal of Cancer

SN - 0007-0920

IS - 3

ER -