Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays.
Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays.
复制标题
使用悬挂滴阵列进行临床前药物敏感性测定，形成稳定的小细胞数三维卵巢癌球体。
DOI：
10.1016/j.ygyno.2015.04.014
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发表时间：
2015-07
影响因子：
4.7
通讯作者：
Mehta G
中科院分区：
文献类型：
--
作者：
Raghavan S;Ward MR;Rowley KR;Wold RM;Takayama S;Buckanovich RJ;Mehta G
卵巢癌从腹水中的多细胞球状聚集体生长和转移。因此，多细胞肿瘤球体是具有生理意义的卵巢癌研究体外模型。传统的悬滴培养需要大量的起始细胞，并且长期维护很繁琐。在这项研究中，我们在新型 384 孔悬滴阵列平台中使用极少量的卵巢癌细胞生成稳定、均匀的多细胞球体。我们使用新型肿瘤球体平台和两种卵巢癌细胞系（A2780 和 OVCAR3）来证明将少至 10 个细胞稳定掺入单个球体中。球体具有均匀的几何形状，投影面积（A2780 球体为 42.60 × 103 μm–475.22 × 103 μm2，OVCAR3 球体为 37.24 × 103 μm2–281.01 × 103 μm2）随初始细胞接种密度而变化。鬼笔环肽和核染色表明细胞形成紧密堆积的球体，具有明确的边界和球体内细胞与细胞的相互作用。球体内的细胞表现出超过 85% 的活力。与 2D 培养物（30-50% 存活率）相比，3D 肿瘤球体对顺铂化疗表现出更大的耐药性（70-80% 存活率）。可以在高通量 384 孔板中用有限的细胞数量生成具有高活力的卵巢癌球状体。与传统二维培养中的细胞相比，球体表现出治疗耐药性。当将此研究转化为稀有的患者来源细胞时，低细胞数量的稳定掺入是有利的。该系统可用于了解卵巢癌球体生物学，以及进行临床前药物敏感性测定。
Ovarian cancer grows and metastasizes from multicellular spheroidal aggregates within the ascites fluid. Multicellular tumor spheroids are therefore physiologically significant3Din vitro models for ovarian cancer research. Conventional hanging drop cultures require high starting cell numbers, and are tedious for long-term maintenance. In this study, we generate stable, uniform multicellular spheroids using very small number of ovarian cancer cells in a novel 384 well hanging drop array platform. We used novel tumor spheroid platform and two ovarian cancer cell lines (A2780 and OVCAR3) to demonstrate the stable incorporation of as few as 10 cells into a single spheroid. Spheroids had uniform geometry, with projected areas (42.60 × 103 μm–475.22 × 103 μm2 for A2780 spheroids and 37.24 × 103 μm2–281.01 × 103 μm2 for OVCAR3 spheroids) that varied as a function of the initial cell seeding density. Phalloidin and nuclear stains indicated cells formed tightly packed spheroids with demarcated boundaries and cell–cell interaction within spheroids. Cells within spheroids demonstrated over 85% viability. 3D tumor spheroids demonstrated greater resistance (70–80% viability) to cisplatin chemotherapy compared to 2D cultures (30–50% viability). Ovarian cancer spheroids can be generated from limited cell numbers in high throughput 384 well plates with high viability. Spheroids demonstrate therapeutic resistance relative to cells in traditional 2D culture. Stable incorporation of low cell numbers is advantageous when translating this research to rare patient-derived cells. This system can be used to understand ovarian cancer spheroid biology, as well as carry out preclinical drug sensitivity assays.
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影响因子：
3.8
作者：
Ferrandina G;Martinelli E;Petrillo M;Prisco MG;Zannoni G;Sioletic S;Scambia G
通讯作者：
Scambia G
影响因子：
24.3
作者：
Correia, Ana Luisa;Bissell, Mina J.
通讯作者：
Bissell, Mina J.
影响因子：
254.7
作者：
Jemal, Ahmedin;Siegel, Rebecca;Thun, Michael J.
通讯作者：
Thun, Michael J.
影响因子：
3.8
作者：
Longati P;Jia X;Eimer J;Wagman A;Witt MR;Rehnmark S;Verbeke C;Toftgård R;Löhr M;Heuchel RL
通讯作者：
Heuchel RL
DOI：
10.1016/j.jconrel.2012.04.045
发表时间：
2012-12-10
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Mehta G;Hsiao AY;Ingram M;Luker GD;Takayama S
通讯作者：
Takayama S