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3D细胞活力检测,需要“极致”手段!

时间:2018-09-27来源:成都溶海华康生物科技有限公司浏览:1110次

然而传统的2D细胞活力检测试剂无法有效的满足3D细胞活力检测需求。



今天我们就来看一看,3D细胞活力检测的“极致”手段。


CellTiter-Glo® 3D 发光法细胞活力检测试剂盒通过对ATP 的定量来测定3D 培养细胞的细胞活力。ATP 是活细胞新陈代谢的指标。CellTiter-Glo® 3D 细胞活力检测试剂的特殊配方使其拥有更强劲的裂解能力,专门用于检测3D 细胞培养产生的微组织(microtissue)的细胞活力,适用于高通量筛选(HTS)。


应用:已成功于多种方式培养的3D 微组织的活力检测,包括细胞悬滴培养板(hanging-drop plate)、超低吸附细胞培养板、Matrigel 包被的细胞培养板、琼脂糖包被细胞板、甲基纤维素覆盖培养液和 Alvetex® 三维细胞培养体系中培养的3D 微组织。

将不同数量的 HCT116 细胞 (RPMI 培养基 +10% FBS) 接种在 InSphero GravityPLUS 96- 孔悬滴培养板, 培养 4 天。 

A. 在微组织培养孔中, 加入与培养基等体积的CellTiter-Glo® 3D 或其他厂家的 A 检测试剂, 震荡混匀 5 分钟后, 室温孵育 30 分钟,应用 GloMax® 发光检测仪读数。 

B. 2X CellTox 绿色荧光染料 ( 结合于膜完整性受损的细胞的双链 DNA) 提前预混在 CellTiter-Glo® 3D 检测试剂 ( 左 ) 或 A 检测试剂 ( 右 ) 中,后共同加入细胞培养孔。 震荡混匀 5 分钟后, 室温孵育 30 分钟, 应用激光共聚焦显微镜拍照。 绿色荧光染色显示裂解的细胞。 微组织直径为 300μm。




03

与不同的3D 细胞培养系统均兼容

HCT116 结肠癌细胞以不同的平台进行培养:hanging-drop 培养400 个细胞,ULA 和Matrigel® matrix 培养1000 个细胞。生长4 天后,微组织以化合物处理48 小时,然后加入等体积CellTiter-Glo® 3D 试剂,震荡5 分钟后,读取数据。

部分应用CTG 3D的已发表文献

1.3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained, Scientific Reports, 2016

 

2.Patterning of tissue spheroids biofabricated from human fibroblasts on the surface of electrospun polyurethane matrix using 3D bioprinter, International Journal of Bioprinting, 2016


3.Inhibition of monocarboxylate transporter 1 suppresses the proliferation of glioblastoma stem cells, The Journal of Physiological Sciences, 2016

 

4.Influence of Matrices on 3D-Cultured Prostate Cancer Cells' Drug Response and Expression of Drug-Action Associated Proteins, Plos one, 2016

 

5.cRGD peptide installation on cisplatin-loaded nanomedicines enhances efficacy against locally advanced head and neck squamous cell carcinoma bearing cancer stem-like cells, Journal of Controlled Release, 2017

 

6.Monitoring the effects of doxorubicin on 3D-spheroid tumor cells in real-time, Onco Targets Ther, 2016

 

7.Prolyl isomerase Pin1 promotes survival in EGFR-mutant lung adenocarcinoma cells with an epithelial–mesenchymal transition phenotype, Laboratory Investigation, 2016

 

8.Drug Discovery Goes Three-Dimensional: Goodbye to Flat High-Throughput Screening? ASSAY and Drug Development Technologies, 2015

 

9.A bioengineered three-dimensional cell culture platform integrated with microfluidics to address antimicrobial resistance in tuberculosis, mBio, 2017

 

10.Impact of adjustable cryogel properties on the performance of prostate cancer cells in 3D, SpringerPlus, 2016

 

11.Silk scaffolds connected with different naturally occurring biomaterials for prostate cancer cell cultivation in 3 D, Biopolymers, 2016

 

12.A High-Content Screening of Anticancer Compounds Suggests the Multiple Tyrosine Kinase Inhibitor Ponatinib for Repurposing in Neuroblastoma Therapy, AACR Journals, 2018

 

13.Overactive mTOR signaling leads to endometrial hyperplasia in aged women and mice, Molecular cancer therapeutics, 2017

 

14.Quantitative high-throughput phenotypic screening of pediatric cancer cell lines identifies multiple opportunities for drug repurposing, Oncotarget, 2018

 

15.Elevation of sensitivity to anticancer agents of human lung adenocarcinoma A549 cells by knockdown of claudin-2 expression in monolayer and spheroid culture models, Biophysica Acta, 2018


16.Miniring approach for high-throughput drug screenings in 3D tumor models, bioRxiv, 2017

 

17.A PDX/organoid biobank of advanced prostate cancers captures genomic and phenotypic heterogeneity for disease modeling and therapeutic screening, Clinical cancer research, 2018

 

18.C3aR and C5aR1 act as key regulators of human and mouse β-cell function, Cellular and Molecular Life Sciences, 2018

 

19.Establishment of patient-derived tumor spheroids for non-small cell lung cancer, PLOS one, 2018

 

20.Evaluation of anticancer agents using patient-derived tumor organoids characteristically similar to source tissues, Oncology Reports, 2018

 

21.DGC-specific RHOA mutations maintained cancer cell survival and promoted cell migration via ROCK inactivation, Oncotarget, 2018

 

22.A novel three-dimensional high-throughput screening approach identifies inducers of a mutant KRAS selective lethal phenotype, Oncogene, 2018

 

23.Cytotoxicity evaluation using cryopreserved primary human hepatocytes in various culture formats, Toxicology letters, 2016

 

24.Endogenous replication stress marks melanomas sensitive to CHK1 inhibitors in vivo, Clinical cancer research , 2018

 

25.Toxicology studies of primycin-sulphate using a three-dimensional (3D) in vitro human liver aggregate model, Toxicology letters, 2017


购买有礼


即日起至2018.10.30购买CellTiter-Glo® 3D 发光法细胞活力检测试剂盒,在后台留言提供详细的姓名、快递地址、电话和购买凭证(包括批次号),即可获赠一个暖绒搭搭毯。



注:封面图片和表情来源网络,侵删。