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Cancer stem cell-like cells derived from malignant peripheral nerve sheath tumors.

Spyra M, Kluwe L, Hagel C, Nguyen R, Panse J, Kurtz A, Mautner VF, Rabkin SD, Demestre M - PLoS ONE (2011)

Bottom Line: Clonal spheres were obtained, which could be passaged multiple times.Furthermore, cells of these clonal S462 spheres differentiated into Schwann cells, smooth muscle/fibroblast and neurons-like cells under specific differentiation-inducing cultural conditions.Finally, subcutaneous injection of the spheres into immunodeficient nude mice led to tumor formation at a higher rate compared to the parental adherent cells (66% versus 10% at 2.5 × 10(5)).

View Article: PubMed Central - PubMed

Affiliation: Department of Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

ABSTRACT
This study aims to examine whether or not cancer stem cells exist in malignant peripheral nerve sheath tumors (MPNST). Cells of established lines, primary cultures and freshly dissected tumors were cultured in serum free conditions supplemented with epidermal and fibroblast growth factors. From one established human MPNST cell line, S462, cells meeting the criteria for cancer stem cells were isolated. Clonal spheres were obtained, which could be passaged multiple times. Enrichment of stem cell-like cells in these spheres was also supported by increased expression of stem cell markers such as CD133, Oct4, Nestin and NGFR, and decreased expression of mature cell markers such as CD90 and NCAM. Furthermore, cells of these clonal S462 spheres differentiated into Schwann cells, smooth muscle/fibroblast and neurons-like cells under specific differentiation-inducing cultural conditions. Finally, subcutaneous injection of the spheres into immunodeficient nude mice led to tumor formation at a higher rate compared to the parental adherent cells (66% versus 10% at 2.5 × 10(5)). These results provide evidence for the existence of cancer stem cell-like cells in malignant peripheral nerve sheath tumors.

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Comparative expression of stem cell markers and mature cell markers in adherent and S462 spheres.(A) Real-Time PCR. Expression of each marker in the spheres was normalized against expression of the same marker in the adherent S462 cells. Spheres at passages between 13 and 16. * p<0.05; adherent versus sphere transcript expression (Nestin, NGFR, Oct4, Sox2, CD133, Sox9, EGFR and NCAM). (B) Flow cytometry. Changes in the percentage of cells expressing mature and stem cell progenitor markers in adherent cells and in spheres (passages 8 and 13), CD133 and NCAM adherent versus spheres p<0.05 *, CD34 and CD90 adherent versus spheres p<0.01 **, NGFR and Nestin adherent versus spheres p<0.01 ***. EGFR, epidermal growth factor receptor; NGFR, nerve growth factor receptor; NCAM, cell adhesion molecule.
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pone-0021099-g003: Comparative expression of stem cell markers and mature cell markers in adherent and S462 spheres.(A) Real-Time PCR. Expression of each marker in the spheres was normalized against expression of the same marker in the adherent S462 cells. Spheres at passages between 13 and 16. * p<0.05; adherent versus sphere transcript expression (Nestin, NGFR, Oct4, Sox2, CD133, Sox9, EGFR and NCAM). (B) Flow cytometry. Changes in the percentage of cells expressing mature and stem cell progenitor markers in adherent cells and in spheres (passages 8 and 13), CD133 and NCAM adherent versus spheres p<0.05 *, CD34 and CD90 adherent versus spheres p<0.01 **, NGFR and Nestin adherent versus spheres p<0.01 ***. EGFR, epidermal growth factor receptor; NGFR, nerve growth factor receptor; NCAM, cell adhesion molecule.

Mentions: Real-Time PCR revealed up-regulation of stem cell markers Nestin, NGFR, Oct4, Notch4, SOX2, CD133 and SOX9 in S462 spheres of passages between 13 and 16, in comparison to S462 adherent cells kept under the SCM culturing conditions for 14–16 hours (Fig. 3A, left). In contrast, EGF receptor (EGFR) and NCAM, a marker of immature Schwann cells, were expressed at lower levels in S462 spheres than in their parental adherent cells (Fig. 3A, right).


Cancer stem cell-like cells derived from malignant peripheral nerve sheath tumors.

Spyra M, Kluwe L, Hagel C, Nguyen R, Panse J, Kurtz A, Mautner VF, Rabkin SD, Demestre M - PLoS ONE (2011)

Comparative expression of stem cell markers and mature cell markers in adherent and S462 spheres.(A) Real-Time PCR. Expression of each marker in the spheres was normalized against expression of the same marker in the adherent S462 cells. Spheres at passages between 13 and 16. * p<0.05; adherent versus sphere transcript expression (Nestin, NGFR, Oct4, Sox2, CD133, Sox9, EGFR and NCAM). (B) Flow cytometry. Changes in the percentage of cells expressing mature and stem cell progenitor markers in adherent cells and in spheres (passages 8 and 13), CD133 and NCAM adherent versus spheres p<0.05 *, CD34 and CD90 adherent versus spheres p<0.01 **, NGFR and Nestin adherent versus spheres p<0.01 ***. EGFR, epidermal growth factor receptor; NGFR, nerve growth factor receptor; NCAM, cell adhesion molecule.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3113907&req=5

pone-0021099-g003: Comparative expression of stem cell markers and mature cell markers in adherent and S462 spheres.(A) Real-Time PCR. Expression of each marker in the spheres was normalized against expression of the same marker in the adherent S462 cells. Spheres at passages between 13 and 16. * p<0.05; adherent versus sphere transcript expression (Nestin, NGFR, Oct4, Sox2, CD133, Sox9, EGFR and NCAM). (B) Flow cytometry. Changes in the percentage of cells expressing mature and stem cell progenitor markers in adherent cells and in spheres (passages 8 and 13), CD133 and NCAM adherent versus spheres p<0.05 *, CD34 and CD90 adherent versus spheres p<0.01 **, NGFR and Nestin adherent versus spheres p<0.01 ***. EGFR, epidermal growth factor receptor; NGFR, nerve growth factor receptor; NCAM, cell adhesion molecule.
Mentions: Real-Time PCR revealed up-regulation of stem cell markers Nestin, NGFR, Oct4, Notch4, SOX2, CD133 and SOX9 in S462 spheres of passages between 13 and 16, in comparison to S462 adherent cells kept under the SCM culturing conditions for 14–16 hours (Fig. 3A, left). In contrast, EGF receptor (EGFR) and NCAM, a marker of immature Schwann cells, were expressed at lower levels in S462 spheres than in their parental adherent cells (Fig. 3A, right).

Bottom Line: Clonal spheres were obtained, which could be passaged multiple times.Furthermore, cells of these clonal S462 spheres differentiated into Schwann cells, smooth muscle/fibroblast and neurons-like cells under specific differentiation-inducing cultural conditions.Finally, subcutaneous injection of the spheres into immunodeficient nude mice led to tumor formation at a higher rate compared to the parental adherent cells (66% versus 10% at 2.5 × 10(5)).

View Article: PubMed Central - PubMed

Affiliation: Department of Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

ABSTRACT
This study aims to examine whether or not cancer stem cells exist in malignant peripheral nerve sheath tumors (MPNST). Cells of established lines, primary cultures and freshly dissected tumors were cultured in serum free conditions supplemented with epidermal and fibroblast growth factors. From one established human MPNST cell line, S462, cells meeting the criteria for cancer stem cells were isolated. Clonal spheres were obtained, which could be passaged multiple times. Enrichment of stem cell-like cells in these spheres was also supported by increased expression of stem cell markers such as CD133, Oct4, Nestin and NGFR, and decreased expression of mature cell markers such as CD90 and NCAM. Furthermore, cells of these clonal S462 spheres differentiated into Schwann cells, smooth muscle/fibroblast and neurons-like cells under specific differentiation-inducing cultural conditions. Finally, subcutaneous injection of the spheres into immunodeficient nude mice led to tumor formation at a higher rate compared to the parental adherent cells (66% versus 10% at 2.5 × 10(5)). These results provide evidence for the existence of cancer stem cell-like cells in malignant peripheral nerve sheath tumors.

Show MeSH
Related in: MedlinePlus