| 潘 岩,花永强,刘鲁明.无血清培养法分离并鉴定胰腺癌细胞系BxPC-3中肿瘤干细胞[J].中国肿瘤,2012,21(6):441-445. |
| 无血清培养法分离并鉴定胰腺癌细胞系BxPC-3中肿瘤干细胞 |
| Isolation and Identification of Pancreatic Cancer Stem Cells from BxPC-3 Cell Line Cultured in Serum-Free Medium |
| 投稿时间：2012-01-03 |
| DOI： |
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| 中文关键词: 胰腺癌 肿瘤干细胞 无血清培养 OCT-4 分离 鉴定 |
| 英文关键词:pancreatic cancer cancer stem cells serum-free medium culture OCT-4 isolation identification |
| 基金项目:国家自然科学基金(81072942;81173461;30901911) |
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| 摘要点击次数: 1748 |
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| 中文摘要: |
| 摘 要：［目的］ 从人胰腺癌细胞系BxPC-3中分离并鉴定胰腺癌干细胞。［方法］用无血清培养基培养人胰腺癌BxPC-3得到肿瘤细胞球。将肿瘤细胞球传代扩增，观察各传代后细胞成球能力；并用含血清培养基诱导促使其分化；蛋白印迹法测定肿瘤干细胞特异性转录因子OCT-4蛋白表达情况，并将肿瘤细胞球细胞植入NOD/SCID小鼠皮下，观察移植瘤的形成。［结果］在无血清培养基中，BxPC-3细胞能形成少量肿瘤细胞球，具有很强的自我更新能力，成球能力随着球体细胞传代次数的增加而增加。在含血清环境中，球体细胞逐渐分化而呈贴壁生长。肿瘤细胞球OCT-4蛋白表达明显高于BxPC-3贴壁细胞。1×104个球体细胞即能在NOD/SCID小鼠皮下成瘤。［结论］无血清培养能够有效的分离和扩增人胰腺癌细胞系BxPC-3的肿瘤干细胞。 |
| 英文摘要: |
| Abstract: ［Purpose］ To isolate and identify pancreatic cancer stem cells from human pancreatic cancer cell line BxPC-3. ［Method］BxPC-3 cells were cultured in serum-free medium to derive tumor spheres. Tumor spheres were then expanded and passage to estimate the potency of sphere-forming，then cultured in serum-containing medium to induce their differentiation. The protein expression of stem cell related transcript factor OCT-4 was evaluated by western blot. The tumorigenesis of the sphere-forming cells was observed in subcutaneously implanted NOD/SCID mice models. ［Results］ A few of floating tumor spheres were isolated and expanded in serum-free medium. These tumor spheres attached to the bottom of culture plates and began to differentiate when cultured in serum-containing medium. The protein expression of OCT-4 was significantly higher in sphere-forming cells of BxPC-3 than that attached cells. Only one thousand spheroid cells can form tumor in NOD/SCID mice. ［Conclusion］ Serum-free medium culture is an effective way to isolate and expand the pancreatic cancer stem cell from BxPC-3. |
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