CAS OpenIR  > 中科院上海应用物理研究所2011-2020年
Encapsulation and release of living tumor cells using hydrogels with the hybridization chain reaction
Ye, DK; Li, M; Zhai, TT; Song, P; Song, L; Wang, H; Mao, XH; Wang, F; Zhang, XL; Ge, ZL; Shi, JY; Wang, LH; Fan, CH; Li, Q; Zuo, XL
2020
Source PublicationNATURE PROTOCOLS
ISSN1754-2189
Volume15Issue:7Pages:2163-2185
Subtype期刊论文
AbstractCirculating tumor cells (CTCs) enable noninvasive liquid biopsy and identification of cancer. Various approaches exist for the capture and release of CTCs, including microfluidic methods and those involving magnetic beads or nanostructured solid interfaces. However, the concomitant cell damage and fragmentation that often occur during capture make it difficult to extensively characterize and analyze living CTCs. Here, we describe an aptamer-trigger-clamped hybridization chain reaction (atcHCR) method for the capture of CTCs by porous 3D DNA hydrogels. The 3D environment of the DNA networks minimizes cell damage, and the CTCs can subsequently be released for live-cell analysis. In this protocol, initiator DNAs with aptamer-toehold biblocks specifically bind to the epithelial cell adhesion molecule (EpCAM) on the surface of CTCs, which triggers the atcHCR and the formation of a DNA hydrogel. The DNA hydrogel cloaks the CTCs, facilitating quantification with minimal cell damage. This method can be used to quantitively identify as few as 10 MCF-7 cells in a 2-mu L blood sample. Decloaking of tumor cells via gentle chemical stimulus (ATP) is used to release living tumor cells for subsequent cell culture and live-cell analysis. We also describe how to use the protocol to encapsulate and release cells of cancer cell lines, which can be used in preliminary experiments to model CTCs. The whole protocol takes similar to 2.5 d to complete, including downstream cell culture and analysis.
KeywordDNA HYDROGEL BIOSENSING INTERFACE CAPTURE AMPLIFICATION RECOGNITION PLATFORM
DOI10.1038/s41596-020-0326-4
Indexed BySCI
Language英语
Citation statistics
Cited Times:12[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/32848
Collection中科院上海应用物理研究所2011-2020年
Affiliation1.Shanghai Jiao Tong Univ, Sch Med, Renji Hosp, Inst Mol Med,Dept Lab Med,Shanghai Key Lab Nucle, Shanghai, Peoples R China
2.Shanghai Jiao Tong Univ, Sch Chem & Chem Engn, Shanghai, Peoples R China
3.Chinese Acad Sci, Shanghai Inst Appl Phys, Div Phys Biol, Shanghai Synchrotron Radiat Facil SSRF,CAS Key La, Shanghai, Peoples R China
4.Chinese Acad Sci, Shanghai Inst Appl Phys, Bioimaging Ctr, Shanghai Synchrotron Radiat Facil SSRF,CAS Key La, Shanghai, Peoples R China
5.Shanghai Jiao Tong Univ, Joint Res Ctr Precis Med, Shanghai, Peoples R China
6.Southern Med Univ, Affiliated Fengxian Hosp, Affiliated Sixth Peoples Hosp, South Campus, Shanghai, Peoples R China
7.Chinese Acad Sci, Shanghai Adv Res Inst, Zhangjiang Lab, Shanghai Synchrotron Radiat Facil, Shanghai, Peoples R China
Recommended Citation
GB/T 7714
Ye, DK,Li, M,Zhai, TT,et al. Encapsulation and release of living tumor cells using hydrogels with the hybridization chain reaction[J]. NATURE PROTOCOLS,2020,15(7):2163-2185.
APA Ye, DK.,Li, M.,Zhai, TT.,Song, P.,Song, L.,...&Zuo, XL.(2020).Encapsulation and release of living tumor cells using hydrogels with the hybridization chain reaction.NATURE PROTOCOLS,15(7),2163-2185.
MLA Ye, DK,et al."Encapsulation and release of living tumor cells using hydrogels with the hybridization chain reaction".NATURE PROTOCOLS 15.7(2020):2163-2185.
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