点评详情
发布于:2018-11-9 03:29:43  访问:46 次 回复:0 篇
版主管理 | 推荐 | 删除 | 删除并扣分
Ass substrate was employed to recover cancer cells spiked in blood
Herein we introduce our new microfluidic CTC capture platform and demonstrate its utility in recovering JNK-IN-8 order cancer cells with heterogeneous molecular properties and those obtained from two mouse models of cancer. Our microfluidic CTC capture platform integrates two functional components: 1) a RIE-generated nanoroughened glass substrate with nanoscale topological structures to improve adherent interactions among the glass substrate and cancer cells, and 2) an overlaid polydimethylsiloxane (PDMS) chip using a low profile microfluidic capture chamber that promotes CTC-substrate speak to frequency. Within this work we showed that the microfluidic CTC capture chip could capture > 80 ofChen et al. BMC Cancer (2016) 16:Web page three ofbreast and lung cancer cells spiked in whole blood samples independent in the cell lines‘ EpCAM expression. The microfluidic CTC capture chip also captured equally properly A549 lung cancer cells in their epithelial- or mesenchymal-like state ahead of and after transforming development factor beta (TGF-)-induced EMT. To additional demonstrate the clinical utility from the microfluidic CTC capture chip, we collected complete blood from mice with breast cancer orthotopic xenografts and demonstrated outstanding label-free CTC capture efficiency by the microfluidic CTC capture chip. A lot more importantly, in a syngeneic mouse model of lung cancer using cell lines with known metastatic and non-metastatic capabilities, CTCs were detected in each of the mice with a detectable main tumor independent of the metastatic propensity of the cell line implanted. This highlights the truth that not all CTCs are capable of forming and proliferating as metastases and our newly created microfluidic CTC capture device is able to recover this much less metastatically potent population too.chamber was generated by replica molding utilizing a Si mold fabricated employing microfabrication. The detailed protocol for fabrication from the microfluidic CTC capture chip is described inside the Additional file 1.Human blood specimensHuman blood specimens from wholesome donors were collected in EDTA-containing vacutainers and were processed and assayed inside 6 h of collection. RBC Lysis Buffer (eBioscience) was added to whole blood PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/27488460 at a ten:1 v/v ratio. Following incubation for ten min at space temperature, the sample was diluted with 20?0 mL PBS to quit the lysing reaction then centrifuged at 300 g for 10 min. After discarding the supernatant, the cell pellet was Decernotinib web re-suspended in an equivalent volume of development medium just before use in CTC capture assays.Mouse models of cancerMethodsCell cultureMCF-7, MDA-MB-231, and A549 cells have been acquired from ATCC and SUM149 cells were certified by means of brief tandem repeat evaluation (FTA barcode STR13871). 344SQ and 393P cell lines were derived from K-ras/p53 mutant mice as described in Gibbons et al. [32, 33]. MCF-7 cells had been maintained in high-glucose DMEM (Invitrogen); MDA-MB-231, 344SQ, and 393P cells in RPMI-1640 (Invitrogen); SUM-149 cells in Ham‘s F-12 w/L-glutamine (Fisher S.Ass substrate was employed to recover cancer cells spiked in blood samples, inside a fixed device setting, with capture efficiencies of more than 90 for distinct cancer cell lines [31]. Expanding on this proof-of-concept function, we hypothesized that further improvements in CTC capture functionality and blood sample throughput might be accomplished by using a confining PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28607003 microfluidic environment about the nanoroughened glass substrate to promote cell-substrate interactions for highly efficient CTC capture.
共0篇回复 每页10篇 页次:1/1
共0篇回复 每页10篇 页次:1/1
我要回复
回复内容
验 证 码
看不清?更换一张
匿名发表 
当前位置
脚注信息

美博梁sir2010-2017版权所有