In vitro analysis of breast cancer cell line tumourspheres and primary human breast epithelia mammospheres demonstrates inter- and intrasphere heterogeneity.
In vitro analysis of breast cancer cell line tumourspheres and primary human breast epithelia mammospheres demonstrates inter- and intrasphere heterogeneity.
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DOI：
10.1371/journal.pone.0064388
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Lopez JA
中科院分区：
文献类型：
--
作者：
Smart CE;Morrison BJ;Saunus JM;Vargas AC;Keith P;Reid L;Wockner L;Askarian-Amiri M;Sarkar D;Simpson PT;Clarke C;Schmidt CW;Reynolds BA;Lakhani SR;Lopez JA
乳腺球和乳腺肿瘤球培养作为用于繁殖和分析正常干细胞和癌症干细胞的体外测定而受到欢迎。与其他培养形式相比，来自不同来源或亲代培养物本身的球体是否确实是富含干/祖细胞的单一实体尚未完全确定。我们通过免疫组织化学和流式细胞术调查了 26 个乳腺细胞系、来自 6 个管腔样和基底样细胞系的免疫表型球体的球体形成能力，并使用体外功能测定比较了球体、贴壁和基质胶培养形式之间的克隆形成性。分析揭示了球内和球间的形态和分子异质性，与贴壁亲代细胞系表型一致。流式细胞术显示球体培养物并不普遍富集先前与干细胞表型相关的标记，尽管我们发现球体和贴壁形式之间存在一些细胞系特异性变化。球体形成效率显着低于贴壁或基质胶克隆形成率，并且在连续传代中保持恒定。令人惊讶的是，球体来源的细胞的自我更新能力与其他培养形式相似/较低。我们观察到球体和贴壁培养物中长期增殖细胞对称分裂率之间存在显着相关性，表明维持它们的区室之间存在功能重叠。使用正常原代人乳腺上皮细胞（包括分选的管腔（MUC1+）和基底/肌上皮（CD10+）细胞）进行的实验揭示了原代乳腺球中不同的管腔样、基底样和间质实体。形态学和集落形成细胞测定数据表明，乳腺球培养物可能会富集管腔祖细胞表型，或诱导贴壁培养发生的基础/间质体外选择的逆转/松弛。总体而言，细胞系肿瘤球和原代乳腺球不是富含干细胞的同质实体，这表明需要采取更谨慎的方法来解释这些测定的数据，并仔细考虑其局限性。球体培养可能代表了另一种三维培养系统，该系统不是普遍“富集”干细胞，而是可作为提供祖细胞活性读数的一套功能测定之一。
Mammosphere and breast tumoursphere culture have gained popularity as in vitro assays for propagating and analysing normal and cancer stem cells. Whether the spheres derived from different sources or parent cultures themselves are indeed single entities enriched in stem/progenitor cells compared to other culture formats has not been fully determined. We surveyed sphere-forming capacity across 26 breast cell lines, immunophenotyped spheres from six luminal- and basal-like lines by immunohistochemistry and flow cytometry and compared clonogenicity between sphere, adherent and matrigel culture formats using in vitro functional assays. Analyses revealed morphological and molecular intra- and inter-sphere heterogeneity, consistent with adherent parental cell line phenotypes. Flow cytometry showed sphere culture does not universally enrich for markers previously associated with stem cell phenotypes, although we found some cell-line specific changes between sphere and adherent formats. Sphere-forming efficiency was significantly lower than adherent or matrigel clonogenicity and constant over serial passage. Surprisingly, self-renewal capacity of sphere-derived cells was similar/lower than other culture formats. We observed significant correlation between long-term-proliferating-cell symmetric division rates in sphere and adherent cultures, suggesting functional overlap between the compartments sustaining them. Experiments with normal primary human mammary epithelia, including sorted luminal (MUC1+) and basal/myoepithelial (CD10+) cells revealed distinct luminal-like, basal-like and mesenchymal entities amongst primary mammospheres. Morphological and colony-forming-cell assay data suggested mammosphere culture may enrich for a luminal progenitor phenotype, or induce reversion/relaxation of the basal/mesenchymal in vitro selection occurring with adherent culture. Overall, cell line tumourspheres and primary mammospheres are not homogenous entities enriched for stem cells, suggesting a more cautious approach to interpreting data from these assays and careful consideration of its limitations. Sphere culture may represent an alternative 3-dimensional culture system which rather than universally ‘enriching’ for stem cells, has utility as one of a suite of functional assays that provide a read-out of progenitor activity.
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影响因子：
10.5
作者：
Dontu, G;Abdallah, WM;Wicha, MS
通讯作者：
Wicha, MS
影响因子：
7.4
作者：
Fillmore, Christine M.;Kuperwasser, Charlotte
通讯作者：
Kuperwasser, Charlotte
影响因子：
11.2
作者：
Charafe-Jauffret E;Ginestier C;Iovino F;Wicinski J;Cervera N;Finetti P;Hur MH;Diebel ME;Monville F;Dutcher J;Brown M;Viens P;Xerri L;Bertucci F;Stassi G;Dontu G;Birnbaum D;Wicha MS
通讯作者：
Wicha MS
影响因子：
11.2
作者：
Kern, Scott E.;Shibata, Darryl
通讯作者：
Shibata, Darryl
DOI：
10.1186/bcr2106
发表时间：
2008
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Grimshaw MJ;Cooper L;Papazisis K;Coleman JA;Bohnenkamp HR;Chiapero-Stanke L;Taylor-Papadimitriou J;Burchell JM
通讯作者：
Burchell JM