Limits...
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.

Show MeSH

Related in: MedlinePlus

Proliferation and self-renewal of MPNST spheres.Bromodeoxyuridine incorporation assay revealed proliferation of S462 cells in the clonal spheres (secondary spheres). The increase in cell absorbance was significant for 8, 10, and 15 days in SCM versus 3 days (p<0.01 **). Changes in the frequency of sphere formation in wells containing single cells originally from adherent S462, then from dissociated spheres plated as single cells and passaged consecutively. The increase in frequency of sphere formation with increasing passage was significant (passage 1 (primary spheres) versus passages 10 (10th spheres) and 12 (12th spheres), P<0.01 **, passage 2 (secondary spheres) versus passages 10 (10th spheres) and 12th spheres), P<0.05*).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3113907&req=5

pone-0021099-g002: Proliferation and self-renewal of MPNST spheres.Bromodeoxyuridine incorporation assay revealed proliferation of S462 cells in the clonal spheres (secondary spheres). The increase in cell absorbance was significant for 8, 10, and 15 days in SCM versus 3 days (p<0.01 **). Changes in the frequency of sphere formation in wells containing single cells originally from adherent S462, then from dissociated spheres plated as single cells and passaged consecutively. The increase in frequency of sphere formation with increasing passage was significant (passage 1 (primary spheres) versus passages 10 (10th spheres) and 12 (12th spheres), P<0.01 **, passage 2 (secondary spheres) versus passages 10 (10th spheres) and 12th spheres), P<0.05*).

Mentions: Parental MPNST S462 cells of passage 35, which under standard medium conditions with serum grow adherently, formed floating spheres after 2 days under stem cell conditions (Fig. 1A,B; adherent S462 cells refer to the parental S462 cell line). Proliferation of cells in the spheres demonstrated that these were not just aggregates (Fig. 2A). When these primary spheres were dissociated and the cells plated at extremely low density in wells of a 96-well plate, secondary spheres were obtained in 16.7%±2.4 of wells containing single cells. The frequency of sphere formation from single cells increased when the spheres were passaged further (Fig. 2B). Clonal spheres were also obtained from adherent S462 cells of the very early passage 2, with a higher frequency of primary sphere formation 31.86%±2.50, almost twice as high as with adherent S462 cells of passage 35. To date, MPNST S462 spheres have been passaged under clonal conditions over 20 times.


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)

Proliferation and self-renewal of MPNST spheres.Bromodeoxyuridine incorporation assay revealed proliferation of S462 cells in the clonal spheres (secondary spheres). The increase in cell absorbance was significant for 8, 10, and 15 days in SCM versus 3 days (p<0.01 **). Changes in the frequency of sphere formation in wells containing single cells originally from adherent S462, then from dissociated spheres plated as single cells and passaged consecutively. The increase in frequency of sphere formation with increasing passage was significant (passage 1 (primary spheres) versus passages 10 (10th spheres) and 12 (12th spheres), P<0.01 **, passage 2 (secondary spheres) versus passages 10 (10th spheres) and 12th spheres), P<0.05*).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0021099-g002: Proliferation and self-renewal of MPNST spheres.Bromodeoxyuridine incorporation assay revealed proliferation of S462 cells in the clonal spheres (secondary spheres). The increase in cell absorbance was significant for 8, 10, and 15 days in SCM versus 3 days (p<0.01 **). Changes in the frequency of sphere formation in wells containing single cells originally from adherent S462, then from dissociated spheres plated as single cells and passaged consecutively. The increase in frequency of sphere formation with increasing passage was significant (passage 1 (primary spheres) versus passages 10 (10th spheres) and 12 (12th spheres), P<0.01 **, passage 2 (secondary spheres) versus passages 10 (10th spheres) and 12th spheres), P<0.05*).
Mentions: Parental MPNST S462 cells of passage 35, which under standard medium conditions with serum grow adherently, formed floating spheres after 2 days under stem cell conditions (Fig. 1A,B; adherent S462 cells refer to the parental S462 cell line). Proliferation of cells in the spheres demonstrated that these were not just aggregates (Fig. 2A). When these primary spheres were dissociated and the cells plated at extremely low density in wells of a 96-well plate, secondary spheres were obtained in 16.7%±2.4 of wells containing single cells. The frequency of sphere formation from single cells increased when the spheres were passaged further (Fig. 2B). Clonal spheres were also obtained from adherent S462 cells of the very early passage 2, with a higher frequency of primary sphere formation 31.86%±2.50, almost twice as high as with adherent S462 cells of passage 35. To date, MPNST S462 spheres have been passaged under clonal conditions over 20 times.

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