Expansion of CD133(+) colon cancer cultures retaining stem cell properties to enable cancer stem cell target discovery.
Expansion of CD133(+) colon cancer cultures retaining stem cell properties to enable cancer stem cell target discovery.
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DOI：
10.1038/sj.bjc.6605610
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发表时间：
2010-04-13
影响因子：
8.8
通讯作者：
中科院分区：
文献类型：
--
作者：
尽管早期的研究表明，在体外增殖的实体肿瘤癌干细胞（CSC）作为非粘附肿瘤球，它仍然是有争议的，是否可以在体外维持CSC。肿瘤球的CSC性质的额外验证将支持其用作CSC模型，并提供发现额外CSC细胞表面标志物以帮助CSC检测和潜在消除的机会。从13个手术切除的结肠肿瘤标本中分离的原代肿瘤细胞使用无血清CSC选择性条件繁殖。建立了长期培养的肿瘤球的CSC特性，并进行了基于质谱的蛋白质组学研究。新鲜分离的CD 133+结直肠癌细胞产生了长期的肿瘤球（或球状体）培养物，维持CD 133表达。这些球状体细胞能够自我更新并分化成粘附的上皮细胞系，并重现原始肿瘤的表型。相对于其分化的后代，肿瘤球状体细胞对化疗药物伊立替康更耐药。最后，通过质谱鉴定，CD44、CD166、CD29、CEACAM5、钙粘蛋白17和双糖链蛋白聚糖在CD133+肿瘤球状体细胞中富集。我们的数据表明，从临床标本中分离的离体扩增的结肠CSC可以保持在培养物中，从而能够鉴定CSC细胞表面相关蛋白。
Despite earlier studies demonstrating in vitro propagation of solid tumour cancer stem cells (CSCs) as non-adherent tumour spheres, it remains controversial as to whether CSCs can be maintained in vitro. Additional validation of the CSC properties of tumour spheres would support their use as CSC models and provide an opportunity to discover additional CSC cell surface markers to aid in CSC detection and potential elimination. Primary tumour cells isolated from 13 surgically resected colon tumour specimens were propagated using serum-free CSC-selective conditions. The CSC properties of long-term cultured tumour spheres were established and mass spectrometry-based proteomics performed. Freshly isolated CD133+ colorectal cancer cells gave rise to long-term tumour sphere (or spheroids) cultures maintaining CD133 expression. These spheroid cells were able to self-renew and differentiate into adherent epithelial lineages and recapitulate the phenotype of the original tumour. Relative to their differentiated progeny, tumour spheroid cells were more resistant to the chemotherapeutic irinotecan. Finally, CD44, CD166, CD29, CEACAM5, cadherin 17, and biglycan were identified by mass spectrometry to be enriched in CD133+ tumour spheroid cells. Our data suggest that ex vivo-expanded colon CSCs isolated from clinical specimens can be maintained in culture enabling the identification of CSC cell surface-associated proteins.
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影响因子：
29.4
作者：
Hinoi, T;Lucas, PC;Fearon, ER
通讯作者：
Fearon, ER
影响因子：
64.8
作者：
O'Brien, Catherine A.;Pollett, Aaron;Dick, John E.
通讯作者：
Dick, John E.
影响因子：
12.4
作者：
Eramo, A.;Lotti, F.;De Maria, R.
通讯作者：
De Maria, R.
DOI：
10.1073/pnas.0703478104
发表时间：
2007-06-12
影响因子：
11.1
作者：
Dalerba, Piero;Dylla, Scott J.;Clarke, Michael F.
通讯作者：
Clarke, Michael F.
影响因子：
3.1
作者：
Nishimura, S;Wakabayashi, N;Mitsufuji, S
通讯作者：
Mitsufuji, S