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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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Tumor formation in immunodeficient nude mice.(A) Frequency and time of tumor formation with 2.5×105 adherent (n = 10, passage 35) or clonal sphere S462 (n = 9), CD133+ (n = 9) and CD133− (n = 5) cells injected subcutaneously. All sphere cells were obtained between passages 8 and 13. Adherent versus spheres and adherent versus CD133+ spheres p<0.05 *. (B) Histology of tumors derived from adherent cells (left panel) and sphere cells showed that these tumors resemble MPNSTs. H&E shows typical spindle shape cells of MPNST (arrows, top panel), which are highly proliferative by Ki-67 immunstaining (arrows, middle panel) and negative for S100 (bottom panel). Bar = 200 µm, applies for all the micrographs. (C) Cells dissociated from mice tumors formed secondary spheres in vitro (representative secondary sphere 3 weeks after plating single cells). Bar = 20 µm.
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pone-0021099-g006: Tumor formation in immunodeficient nude mice.(A) Frequency and time of tumor formation with 2.5×105 adherent (n = 10, passage 35) or clonal sphere S462 (n = 9), CD133+ (n = 9) and CD133− (n = 5) cells injected subcutaneously. All sphere cells were obtained between passages 8 and 13. Adherent versus spheres and adherent versus CD133+ spheres p<0.05 *. (B) Histology of tumors derived from adherent cells (left panel) and sphere cells showed that these tumors resemble MPNSTs. H&E shows typical spindle shape cells of MPNST (arrows, top panel), which are highly proliferative by Ki-67 immunstaining (arrows, middle panel) and negative for S100 (bottom panel). Bar = 200 µm, applies for all the micrographs. (C) Cells dissociated from mice tumors formed secondary spheres in vitro (representative secondary sphere 3 weeks after plating single cells). Bar = 20 µm.

Mentions: Subcutaneous injection of 2.5×105 cells of clonal S462 spheres (n = 9) led to visible solid tumor formation 1 to 3 months after injection in 66% of nude mice. In contrast, detectable solid tumors only formed in 10% of nude mice (n = 10) when the same number of adherent S462 cells was injected, this tumor needed over three months to form (Fig. 6A). Histologically, tumors from both adherent cells and spheres exhibited typical characteristics of MPNST: spindle shaped, highly proliferative as shown with high numbers of Ki67 positive cells, and S100 negative tumor cells (Figure 6B).


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)

Tumor formation in immunodeficient nude mice.(A) Frequency and time of tumor formation with 2.5×105 adherent (n = 10, passage 35) or clonal sphere S462 (n = 9), CD133+ (n = 9) and CD133− (n = 5) cells injected subcutaneously. All sphere cells were obtained between passages 8 and 13. Adherent versus spheres and adherent versus CD133+ spheres p<0.05 *. (B) Histology of tumors derived from adherent cells (left panel) and sphere cells showed that these tumors resemble MPNSTs. H&E shows typical spindle shape cells of MPNST (arrows, top panel), which are highly proliferative by Ki-67 immunstaining (arrows, middle panel) and negative for S100 (bottom panel). Bar = 200 µm, applies for all the micrographs. (C) Cells dissociated from mice tumors formed secondary spheres in vitro (representative secondary sphere 3 weeks after plating single cells). Bar = 20 µm.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0021099-g006: Tumor formation in immunodeficient nude mice.(A) Frequency and time of tumor formation with 2.5×105 adherent (n = 10, passage 35) or clonal sphere S462 (n = 9), CD133+ (n = 9) and CD133− (n = 5) cells injected subcutaneously. All sphere cells were obtained between passages 8 and 13. Adherent versus spheres and adherent versus CD133+ spheres p<0.05 *. (B) Histology of tumors derived from adherent cells (left panel) and sphere cells showed that these tumors resemble MPNSTs. H&E shows typical spindle shape cells of MPNST (arrows, top panel), which are highly proliferative by Ki-67 immunstaining (arrows, middle panel) and negative for S100 (bottom panel). Bar = 200 µm, applies for all the micrographs. (C) Cells dissociated from mice tumors formed secondary spheres in vitro (representative secondary sphere 3 weeks after plating single cells). Bar = 20 µm.
Mentions: Subcutaneous injection of 2.5×105 cells of clonal S462 spheres (n = 9) led to visible solid tumor formation 1 to 3 months after injection in 66% of nude mice. In contrast, detectable solid tumors only formed in 10% of nude mice (n = 10) when the same number of adherent S462 cells was injected, this tumor needed over three months to form (Fig. 6A). Histologically, tumors from both adherent cells and spheres exhibited typical characteristics of MPNST: spindle shaped, highly proliferative as shown with high numbers of Ki67 positive cells, and S100 negative tumor cells (Figure 6B).

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