Identification of In-droplet Multicellular Communities by Light-induced Combinatorial DNA Barcoding

Microdroplet co-culture system is useful for the parallel assessment of numerous possible cell-cell interactions by generating isolated subcommunities from a pool of heterogeneous cells. However, the integration of single-cell sequencing into such analysis has been limited due to the lack of effecti...

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Bibliographic Details
Published in:Chemistry : a European journal 2023-09, Vol.29 (53), p.e202301133-e202301133
Main Authors: Kawasaki, Fumiko, Mori, Yuka, Mimori, Takahiro, Sato, Issei, Ota, Sadao
Format: Article
Language:English
Subjects:
Cell culture
Cell interactions
Cell membranes
Chemistry
Combinatorial analysis
Deoxyribonucleic acid
DNA
DNA barcoding
Droplets
Gene sequencing
Microparticles
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Summary:Microdroplet co-culture system is useful for the parallel assessment of numerous possible cell-cell interactions by generating isolated subcommunities from a pool of heterogeneous cells. However, the integration of single-cell sequencing into such analysis has been limited due to the lack of effective molecular identifiers for each in-droplet subcommunity. Here, we present a strategy for generating in-droplet subcommunity identifiers using DNA-functionalized microparticles encapsulated within microdroplets. These microparticles serve as initial information carriers, where their combinations act as distinct identifiers for in-droplet subcommunity. Upon optical trigger, DNA barcoding molecules encoding the microparticle information are once released in the microdroplets and then tag cell membranes. The tagged DNA molecules then serves as a second information carrier readable by single-cell sequencing to reconstitute the community in silico in single-cell RNA sequencing data space.
ISSN:0947-6539
1521-3765
DOI:10.1002/chem.202301133