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.
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: F
authors.initials: F
authors.lastname: Fan
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: S
authors.initials: S
authors.lastname: Bellister
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: J
authors.initials: J
authors.lastname: Lu
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: X
authors.initials: X
authors.lastname: Ye
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: D R
authors.initials: DR
authors.lastname: Boulbes
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: F
authors.initials: F
authors.lastname: Tozzi
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: E
authors.initials: E
authors.lastname: Sceusi
authors.affiliation: Department of Gastrointestinal Medical Oncology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
authors.forename: S
authors.initials: S
authors.lastname: Kopetz
authors.affiliation: Department of Gastrointestinal Medical Oncology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
authors.forename: F
authors.initials: F
authors.lastname: Tian
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: L
authors.initials: L
authors.lastname: Xia
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA.
authors.forename: R
authors.initials: R
authors.lastname: Bhattacharya
authors.affiliation: 1] Department of Surgical Oncology, Unit 1484, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77230-1402, USA [2] Department of Molecular and Cellular Oncology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
authors.forename: L M
authors.initials: LM
authors.lastname: Ellis
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D018920:Coculture Techniques; D015179:Colorectal Neoplasms / Q000473:pathology*; D045325:HCT116 Cells; D019073:HT29 Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000473:pathology* / Q000502:physiology; D061251:Primary Cell Culture / Q000379:methods*; D018874:Spheroids, Cellular / Q000378:metabolism / Q000473:pathology; D014407:Tumor Cells, Cultured
chemical_list: D018960:Hyaluronan Receptors