J Korean Med Sci.  2010 May;25(5):798-803. 10.3346/jkms.2010.25.5.798.

Primary Culture of Central Neurocytoma: A Case Report

• Paek SH
• Shin HY
• Kim JW
• Park SH
• Son JH
• Kim DG

• KMID: 1713970
• DOI: http://doi.org/10.3346/jkms.2010.25.5.798

Abstract

A seventeen-year-old female patient was admitted with sudden-onset of headache and vomiting. Brain magnetic resonance imaging demonstrated a heterogeneously enhancing tumour in the left lateral ventricle. The tumour was removed and confirmed as a central neurocytoma (CN). For the residual tumour in the left lateral ventricle, gamma knife stereotactic radiosurgery was done at fifteen months after the initial surgery. Tumour recurred in the 4th ventricle at 5 yr after initial surgery. The tumour was removed and proved as a CN. In vitro primary culture was done with both tumours obtained from the left lateral ventricle and the 4th ventricle, respectively. Nestin, a neuronal stem cell marker was expressed in reverse Transcriptase-Polymerase Chain Reaction of both tumors. Both tumours showed different morphology and phenotypes of neuron and glia depending on the culture condition. When cultured in insulin, transferrin selenium and fibronectin media with basic fibroblast growth factors, tumour cells showed neuronal morphology and phenotypes. When cultured in the Dulbeco's Modified Essential Media with 20% fetal bovine serum, tumors cells showed glial morphology and phenotypes. It is suggested that CN has the characteristics of neuronal stem cells and potential to differentiate into mature neuron and glial cells depending on the environmental cue.

Keyword

Neurocytoma; Cell Culture Techniques; Neuronal Stem Cells

MeSH Terms

Brain Neoplasms/*pathology
Cell Culture Techniques/methods
Cell Differentiation
Female
Humans
Neurocytoma/*pathology
Stem Cells/*pathology
Young Adult

Figure

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  Fig. 1 Imaging and pathological findings. (A) A T2 weighted brain MRI showed an inhomogeneous mass lesion in the left lateral ventricle. (B) The tumour cells were demonstrated as central neurocytoma (×200). (C) A T1 weighted brain MRI with contrast taken at 15 months after the initial surgery showed a residual mass lesion. (D) A T1 weighted brain MRI with contrast, taken at 45 months after gamma knife radiosurgery showed a ill-defined enhancing mass lesion in the left lateral ventricle (open arrow) and in the 4th ventricle (closed arrow). (E) The tumour in the 4th ventricle was confirmed as the same pathology as the previous one (×200). (F) An electron microscope demonstrated that the perikaryal cytoplasm and cytoplasmic processes have electron dense core neurosecretory granules, 10 nm-neural tubules (white arrow) and a synaptic junction (black arrow), the same finding as in the previous one (×8,000).

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  Fig. 2 RT-PCR of frozen central neurocytoma tumour cells from the initial and second operation demonstrated nestin products. 1, CN from the initial surgery; 2, CN from the second surgery; 3, 1 Kb DNA ladder.

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  Fig. 3 Tumour cells of CN in the left lateral ventricle cultured in 20% DMEM (A-C) and tumour cells of CN in the 4th ventricle cultured in ITSFn with bFGF (D-F). (A) Within several days of primary culture in DMEM with 20% FBS, cultured cells became slightly enlarged and elongated (×200). (B) Several weeks after plating, cultured cells had increased in size with bipolar or multipolar cytoplasmic sprouting (×200). (C) Several months after plating, the cytoplasmic process of cultured cells was markedly enlarged with variable shapes (×400). (D) When cultured in ITSFn with bFGF, uniform small round cultured cells were dominant with their cytoplasmic processes within several days of in vitro primary culture of CN in the 4th ventricle (×200). (E) Several weeks after plating, the round cells had increased in size without elongation of their cytoplasm (×200). (F) Several months after plating, cultured cells displayed round cell morphology without prominent cytoplasmic process (×400).

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  Fig. 4 Morphological changes of tumour cells of CN in the left lateral ventricle cultured in different culture media after thawing; DMEM with 10% FBS (A-C) vs. ITSFn with bFGF (D-F) (×400). (A) Within several days of culture in DMEM with 10% FBS, thawed cells returned to their original morphology at freezing (×400). (B, C) The morphology of the tumour cells at 6 and 20 days after thawing were the same as those in the original cells from the primary culture in DMEM with 20% FBS as shown in Fig. 3B (×400). (D) When cultured in ITSFn with bFGF, the thawed tumour cells returned to their original morphology at freezing within several days (×400). (E, F) The morphology of the tumour cells at 6 and 20 days after thawing were similar to those of primary culture tumour cells cultured in ITSFn with bFGF as shown in Fig. 3E (×400).

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  Fig. 5 Immunohistochemical staining of small round or bipolar tumour cells of CN in the 4th ventricle, cultured in ITSFn with bFGF. Double labeling immunocytochemical studies demonstrated strong positivity for both neuronal and glial markers. Nestin-positive cells were colabeled with Tuj1 (A, ×400), Nestin-positive cells were co-labeled with GFAP (B, ×400). A2B5-positive cells were co-labeled with GFAP (C, ×400), A2B5-positive cells were co-labeled with MAP2 (D, ×400). O4-positive cells were co-labeled with NeuN (E, ×400), GFAP-positive cells were co-labeled with Tuj1 (F, ×400).

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