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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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Multi-lineage differentiation of cells of clonal S462 spheres.Adherent cells (left column) and dissociated sphere cells (right columns) were cultured with growth factors inducing differentiation into cells resembling (A) Schwann cells, (B) SM/Fb and (C) neurons, which are positively stained for S100/NGFR/neurofilament (Schwann cells), SMA (SM/Fb), and MAP-2 (neurons), respectively. Insert in A illustrates S100+ Schwann cells derived from a plexiform neurofibroma culture. (D) phase contrast micrographs from differentiated cells under 3 culture conditions to generate Schwann cell, SM/Fb and neuron-like cells. Differentiated cells are indicated by arrows and non-differentiated cells by arrowheads. Bars = 20 µm. NGFR, nerve growth factor; SMA, smooth muscle actin; MAP-2, microtubule associated protein-2; PNF, plexiform neurofibroma; LC, like-cells.
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pone-0021099-g005: Multi-lineage differentiation of cells of clonal S462 spheres.Adherent cells (left column) and dissociated sphere cells (right columns) were cultured with growth factors inducing differentiation into cells resembling (A) Schwann cells, (B) SM/Fb and (C) neurons, which are positively stained for S100/NGFR/neurofilament (Schwann cells), SMA (SM/Fb), and MAP-2 (neurons), respectively. Insert in A illustrates S100+ Schwann cells derived from a plexiform neurofibroma culture. (D) phase contrast micrographs from differentiated cells under 3 culture conditions to generate Schwann cell, SM/Fb and neuron-like cells. Differentiated cells are indicated by arrows and non-differentiated cells by arrowheads. Bars = 20 µm. NGFR, nerve growth factor; SMA, smooth muscle actin; MAP-2, microtubule associated protein-2; PNF, plexiform neurofibroma; LC, like-cells.

Mentions: Adding neuregulin and forskolin induced specific differentiation to S100+ Schwann cells in nearly all cells of clonal S462 spheres, but only a small portion of adherent S462 cells were really positive for S100 and showed slight different morphology (Fig. 5A). The bi-polar morphology of the S100+ Schwann cells differentiated from clonal spheres (Fig. 5A, D) was similar to that of Schwann cells derived from a plexiform neurofibroma (Fig. 5A insert). These differentiated cells from clonal spheres also expressed the Schwann cell markers neurofilament and NGFR (Fig. 5A), adherent cells expressed neurofilament but very rarely NGFR (data not shown).


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)

Multi-lineage differentiation of cells of clonal S462 spheres.Adherent cells (left column) and dissociated sphere cells (right columns) were cultured with growth factors inducing differentiation into cells resembling (A) Schwann cells, (B) SM/Fb and (C) neurons, which are positively stained for S100/NGFR/neurofilament (Schwann cells), SMA (SM/Fb), and MAP-2 (neurons), respectively. Insert in A illustrates S100+ Schwann cells derived from a plexiform neurofibroma culture. (D) phase contrast micrographs from differentiated cells under 3 culture conditions to generate Schwann cell, SM/Fb and neuron-like cells. Differentiated cells are indicated by arrows and non-differentiated cells by arrowheads. Bars = 20 µm. NGFR, nerve growth factor; SMA, smooth muscle actin; MAP-2, microtubule associated protein-2; PNF, plexiform neurofibroma; LC, like-cells.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0021099-g005: Multi-lineage differentiation of cells of clonal S462 spheres.Adherent cells (left column) and dissociated sphere cells (right columns) were cultured with growth factors inducing differentiation into cells resembling (A) Schwann cells, (B) SM/Fb and (C) neurons, which are positively stained for S100/NGFR/neurofilament (Schwann cells), SMA (SM/Fb), and MAP-2 (neurons), respectively. Insert in A illustrates S100+ Schwann cells derived from a plexiform neurofibroma culture. (D) phase contrast micrographs from differentiated cells under 3 culture conditions to generate Schwann cell, SM/Fb and neuron-like cells. Differentiated cells are indicated by arrows and non-differentiated cells by arrowheads. Bars = 20 µm. NGFR, nerve growth factor; SMA, smooth muscle actin; MAP-2, microtubule associated protein-2; PNF, plexiform neurofibroma; LC, like-cells.
Mentions: Adding neuregulin and forskolin induced specific differentiation to S100+ Schwann cells in nearly all cells of clonal S462 spheres, but only a small portion of adherent S462 cells were really positive for S100 and showed slight different morphology (Fig. 5A). The bi-polar morphology of the S100+ Schwann cells differentiated from clonal spheres (Fig. 5A, D) was similar to that of Schwann cells derived from a plexiform neurofibroma (Fig. 5A insert). These differentiated cells from clonal spheres also expressed the Schwann cell markers neurofilament and NGFR (Fig. 5A), adherent cells expressed neurofilament but very rarely NGFR (data not shown).

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