An Ex Vivo Brain Slice Model to Study and Target Breast Cancer Brain Metastatic Tumor Growth.
An Ex Vivo Brain Slice Model to Study and Target Breast Cancer Brain Metastatic Tumor Growth.
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DOI：
10.3791/62617
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发表时间：
2021-09-22
期刊：
影响因子：
--
通讯作者：
Reginato MJ
中科院分区：
文献类型：
--
作者：
Ciraku L;Moeller RA;Esquea EM;Gocal WA;Hartsough EJ;Simone NL;Jackson JG;Reginato MJ
脑转移是女性乳腺癌的严重后果，因为这些肿瘤难以治疗，并且与不良临床结局相关。乳腺癌脑转移（BCBM）生长的临床前小鼠模型是有用的，但昂贵，并且难以追踪脑实质内的活细胞和肿瘤细胞侵袭。这里提出的是一个协议，离体脑切片培养异种移植小鼠含有颅内注射的乳腺癌脑寻求克隆亚系。将MDA-MB-231 BR荧光素酶标记的细胞颅内注射到Nu/Nu雌性小鼠的脑中，在肿瘤形成后，分离脑，切片并离体培养。对肿瘤切片进行成像以鉴定表达荧光素酶的肿瘤细胞，并监测其在脑实质中的增殖和侵袭长达10天。此外，该方案描述了使用延时显微镜对电离辐射或化疗治疗后肿瘤细胞的生长和侵袭行为进行成像。肿瘤细胞对治疗的反应可以通过活体成像显微镜观察，测量生物发光强度，并在含有BCBM细胞的脑切片上进行组织学检查来观察。因此，这种离体切片模型可以是用于快速测试单独或与辐射组合的新型治疗剂的有用平台，以鉴定个性化的药物，以靶向脑微环境内个体患者的乳腺癌脑转移生长。
Brain metastasis is a serious consequence of breast cancer for women as these tumors are difficult to treat and are associated with poor clinical outcomes. Preclinical mouse models of breast cancer brain metastatic (BCBM) growth are useful but are expensive, and it is difficult to track live cells and tumor cell invasion within the brain parenchyma. Presented here is a protocol for ex vivo brain slice cultures from xenografted mice containing intracranially injected breast cancer brain-seeking clonal sublines. MDA-MB-231BR luciferase tagged cells were injected intracranially into the brains of Nu/Nu female mice, and following tumor formation, the brains were isolated, sliced, and cultured ex vivo. The tumor slices were imaged to identify tumor cells expressing luciferase and monitor their proliferation and invasion in the brain parenchyma for up to 10 days. Further, the protocol describes the use of time-lapse microscopy to image the growth and invasive behavior of the tumor cells following treatment with ionizing radiation or chemotherapy. The response of tumor cells to treatments can be visualized by live-imaging microscopy, measuring bioluminescence intensity, and performing histology on the brain slice containing BCBM cells. Thus, this ex vivo slice model may be a useful platform for rapid testing of novel therapeutic agents alone or in combination with radiation to identify drugs personalized to target an individual patient’s breast cancer brain metastatic growth within the brain microenvironment.
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影响因子：
4
作者：
Fitzgerald, Daniel P.;Palmieri, Diane;Hua, Emily;Hargrave, Elizabeth;Herring, Jeanne M.;Qian, Yongzhen;Vega-Valle, Eleazar;Weil, Robert J.;Stark, Andreas M.;Vortmeyer, Alexander O.;Steeg, Patricia S.
通讯作者：
Steeg, Patricia S.
影响因子：
14
作者：
Fong EL;Harrington DA;Farach-Carson MC;Yu H
通讯作者：
Yu H
影响因子：
4
作者：
Koerfer, Justus;Kallendrusch, Sonja;Lordick, Florian
通讯作者：
Lordick, Florian
影响因子：
3.7
作者：
Mewes A;Franke H;Singer D
通讯作者：
Singer D
DOI：
10.3791/50881
发表时间：
2013-10-09
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Chuang HN;Lohaus R;Hanisch UK;Binder C;Dehghani F;Pukrop T
通讯作者：
Pukrop T