Isolation of Cancer Stem Cells and Astrocytes from Human Glioblastoma: Morphological Characterization of Two Cells Types

M. Hashemi
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Abstract

Abstract  Background: Glioblastoma multiforme is the most aggressive astrocytoma in adults. Glioblastoma is a heterogenous tumor formed from various cells including astrocyte and cancer stem cells. Here, we explain the isolation, culture, morphology and specific markers of human glioblastoma astrocyte and stem cells. Methods: We isolated astrocyte and cancer stem cells from human glioblastoma tissue. The obtained glioblastoma tissues were digested and cultured in DMEM12, B27 supplemented with basic fibroblast growth factor and epidermal growth factor . The morphology and specific markers were assessed in astrocyte and cancer stem cell of human glioblastoma through immunochemistry. Results: Results indicated that there were two morphology types in cell culture including epithelioid morphology and fibroblastic morphology. The astrocyte confirmed via expression of the GFAP protein. Cancer stem cells were round and floating in the culture medium. Immunocytochemical staining indicated that nestin and SOX2 antigens were positively expressed in primary neurospheres. Conclusion: The expression of glial and stem cell markers show that both cells are in the human glioblastoma.
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从人胶质母细胞瘤中分离癌症干细胞和星形胶质细胞:两种细胞类型的形态学特征
摘要背景:多形性胶质母细胞瘤是成人中侵袭性最强的星形细胞瘤。胶质母细胞瘤是一种由星形胶质细胞和癌症干细胞等多种细胞形成的异质性肿瘤。在这里,我们解释了人类胶质母细胞瘤星形胶质细胞和干细胞的分离、培养、形态和特异性标志物。方法:从人胶质母细胞瘤组织中分离星形胶质细胞和癌症干细胞。将获得的胶质母细胞瘤组织在补充有碱性成纤维细胞生长因子和表皮生长因子的DMEM12、B27中消化和培养。用免疫组化方法检测人胶质母细胞瘤星形胶质细胞和癌症干细胞的形态和特异性标志物。结果:细胞培养有上皮样形态和成纤维细胞形态两种形态。星形胶质细胞通过GFAP蛋白的表达得到证实。癌症干细胞呈圆形,漂浮在培养基中。免疫细胞化学染色显示nestin和SOX2抗原在原发性神经球中阳性表达。结论:胶质细胞和干细胞标志物的表达表明这两种细胞都存在于人胶质母细胞瘤中。
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审稿时长
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