Single-Target Pairing System (StarPair) for Large-Scale Interrogation of Cell–Cell Interactions

Tianjiao Mao; Lang Nan; Miao Xu; Kehao Zeng; Yuchao Wang; Ziyu Han; Ho Cheung Shum

doi:10.1002/advs.202513951

Single-Target Pairing System (StarPair) for Large-Scale Interrogation of Cell–Cell Interactions

Tianjiao Mao
, Lang Nan
, Miao Xu
, Kehao Zeng
, Yuchao Wang
, Ziyu Han
, Ho Cheung Shum^*

^*Corresponding author for this work

Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review

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Abstract

Coordination of cell populations through intercellular interactions is essential to the formation and function of living organisms. However, technologies enabling systematic dissection of cell–cell communications or manipulation of cell–cell interactions at the single-cell resolution remain largely out of reach. To solve this, a single-target pairing system, termed StarPair, is proposed, for the high-throughput and high-precision combination of desired single targets in droplets. How target buoyancy, target concentration, sorting parameters, droplet flow rates, and electric stimulation collectively determine the single-target pairing performance is comprehensively explored. Upon optimization of these parameters, pairing efficiencies over 95% can be achieved with an operation frequency reaching one million single-target pairs per 9.5 h for two-target assembly. Leveraging StarPair, the immune cell-cancer cell interactions are assessed between 4 × 10⁵ cell pairs and the NK-92MI cells with high secretion capability are further enriched. It is envisioned that StarPair will open new perspectives for characterizing and manipulating cell–cell interactions at scale. © 2025 The Author(s). Advanced Science published by Wiley-VCH GmbH.

Original language	English
Article number	e13951
Number of pages	14
Journal	Advanced Science
DOIs	https://doi.org/10.1002/advs.202513951
Publication status	Online published - 12 Dec 2025

Funding

The authors acknowledge the financial support provided by the Research Grants Council (RGC) of Hong Kong through the Collaborative Research Fund (C7165-20GF) and, General Research Fund (17303123). Ho Cheung Shum is also supported by the RGC Senior Research Fellow Scheme (SRFS2425-7S04), as well as the Health@InnoHK Program of the Innovation and Technology Commission of the Hong Kong SAR Government. The authors thank Prof. Anskar Yu Hung Leung and his PhD student, Xiaoyuan Zeng, for providing comments and suggestions on manuscript preparation. The authors also thank Dr. Alan Siu Lun Wong for providing comments and suggestions on manuscript revision and Dr. Bei Wang for the assistance with the qPCR experiments and discussions on manuscript revision.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Research Keywords

cell–cell interactions
droplets
functional immune cell screening
single-target pairing

Publisher's Copyright Statement

This full text is made available under CC-BY 4.0. https://creativecommons.org/licenses/by/4.0/

RGC Funding Information

RGC-funded

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10.1002/advs.202513951

396743217.pdfFinal Published version, 2.84 MBLicence: CC BY

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@article{f2c97731ca4d4ab4a7febf0a553698ad,

title = "Single-Target Pairing System (StarPair) for Large-Scale Interrogation of Cell–Cell Interactions",

abstract = "Coordination of cell populations through intercellular interactions is essential to the formation and function of living organisms. However, technologies enabling systematic dissection of cell–cell communications or manipulation of cell–cell interactions at the single-cell resolution remain largely out of reach. To solve this, a single-target pairing system, termed StarPair, is proposed, for the high-throughput and high-precision combination of desired single targets in droplets. How target buoyancy, target concentration, sorting parameters, droplet flow rates, and electric stimulation collectively determine the single-target pairing performance is comprehensively explored. Upon optimization of these parameters, pairing efficiencies over 95\% can be achieved with an operation frequency reaching one million single-target pairs per 9.5 h for two-target assembly. Leveraging StarPair, the immune cell-cancer cell interactions are assessed between 4 × 105 cell pairs and the NK-92MI cells with high secretion capability are further enriched. It is envisioned that StarPair will open new perspectives for characterizing and manipulating cell–cell interactions at scale. {\textcopyright} 2025 The Author(s). Advanced Science published by Wiley-VCH GmbH.",

keywords = "cell–cell interactions, droplets, functional immune cell screening, single-target pairing",

author = "Tianjiao Mao and Lang Nan and Miao Xu and Kehao Zeng and Yuchao Wang and Ziyu Han and Shum, \{Ho Cheung\}",

year = "2025",

month = dec,

day = "12",

doi = "10.1002/advs.202513951",

language = "English",

journal = "Advanced Science",

issn = "2198-3844",

publisher = "Wiley-VCH Verlag GmbH",

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TY - JOUR

T1 - Single-Target Pairing System (StarPair) for Large-Scale Interrogation of Cell–Cell Interactions

AU - Mao, Tianjiao

AU - Nan, Lang

AU - Xu, Miao

AU - Zeng, Kehao

AU - Wang, Yuchao

AU - Han, Ziyu

AU - Shum, Ho Cheung

PY - 2025/12/12

Y1 - 2025/12/12

N2 - Coordination of cell populations through intercellular interactions is essential to the formation and function of living organisms. However, technologies enabling systematic dissection of cell–cell communications or manipulation of cell–cell interactions at the single-cell resolution remain largely out of reach. To solve this, a single-target pairing system, termed StarPair, is proposed, for the high-throughput and high-precision combination of desired single targets in droplets. How target buoyancy, target concentration, sorting parameters, droplet flow rates, and electric stimulation collectively determine the single-target pairing performance is comprehensively explored. Upon optimization of these parameters, pairing efficiencies over 95% can be achieved with an operation frequency reaching one million single-target pairs per 9.5 h for two-target assembly. Leveraging StarPair, the immune cell-cancer cell interactions are assessed between 4 × 105 cell pairs and the NK-92MI cells with high secretion capability are further enriched. It is envisioned that StarPair will open new perspectives for characterizing and manipulating cell–cell interactions at scale. © 2025 The Author(s). Advanced Science published by Wiley-VCH GmbH.

AB - Coordination of cell populations through intercellular interactions is essential to the formation and function of living organisms. However, technologies enabling systematic dissection of cell–cell communications or manipulation of cell–cell interactions at the single-cell resolution remain largely out of reach. To solve this, a single-target pairing system, termed StarPair, is proposed, for the high-throughput and high-precision combination of desired single targets in droplets. How target buoyancy, target concentration, sorting parameters, droplet flow rates, and electric stimulation collectively determine the single-target pairing performance is comprehensively explored. Upon optimization of these parameters, pairing efficiencies over 95% can be achieved with an operation frequency reaching one million single-target pairs per 9.5 h for two-target assembly. Leveraging StarPair, the immune cell-cancer cell interactions are assessed between 4 × 105 cell pairs and the NK-92MI cells with high secretion capability are further enriched. It is envisioned that StarPair will open new perspectives for characterizing and manipulating cell–cell interactions at scale. © 2025 The Author(s). Advanced Science published by Wiley-VCH GmbH.

KW - cell–cell interactions

KW - droplets

KW - functional immune cell screening

KW - single-target pairing

UR - https://www.scopus.com/pages/publications/105024675639

UR - https://www.scopus.com/record/pubmetrics.uri?eid=2-s2.0-105024675639&origin=recordpage

U2 - 10.1002/advs.202513951

DO - 10.1002/advs.202513951

M3 - RGC 21 - Publication in refereed journal

SN - 2198-3844

JO - Advanced Science

JF - Advanced Science

M1 - e13951

ER -

Single-Target Pairing System (StarPair) for Large-Scale Interrogation of Cell–Cell Interactions

Abstract

Funding

UN SDGs

Research Keywords

Publisher's Copyright Statement

RGC Funding Information

Access Output

Other Access Options

Permanent Link

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SRFS: Deciphering Sequence-Dependent Molecular Interactions in Oligonucleotide-Based Coacervates: A Paradigm Shift in Biomaterial Design and Biomedical Engineering

GRF: Single-cell-to-single-cell (SC2SC) Pairing for Hybridoma Production and Monoclonal Antibody Development

Cite this

Single-Target Pairing System (StarPair) for Large-Scale Interrogation of Cell–Cell Interactions

Abstract

Funding

UN SDGs

Research Keywords

Publisher's Copyright Statement

RGC Funding Information

Access Output

Other Access Options

Permanent Link

Other Files and Links

Fingerprint

Projects

SRFS: Deciphering Sequence-Dependent Molecular Interactions in Oligonucleotide-Based Coacervates: A Paradigm Shift in Biomaterial Design and Biomedical Engineering

GRF: Single-cell-to-single-cell (SC2SC) Pairing for Hybridoma Production and Monoclonal Antibody Development

Cite this