Organoid chip based automatic system for long-term quantifying ciliary beating under drug intervention

TY - JOUR

T1 - Organoid chip based automatic system for long-term quantifying ciliary beating under drug intervention

AU - Li, Zixi

AU - Huang, Zhicheng

AU - Wang, Daoyun

AU - Li, Tong

AU - Peng, Junya

AU - Zhang, Anlan

AU - Sun, Ruimeng

AU - Wu, Xin

AU - Zhang, Yu

AU - Liang, Naixin

AU - Li, Qin

AU - Wei, Zewen

N1 - Publisher Copyright: © 2025

PY - 2026/1/22

Y1 - 2026/1/22

N2 - Background: The beating of motile cilia plays an important role in many physiological processes. In recent years, the precise monitoring of ciliary beating has provided valuable insights into cilia-related diseases and their therapies. However, a gap persists between homogeneous in vitro cell models and heterogeneous in vivo ciliary systems. Human-derived ciliary organoids (HDCOs) show promise in bridging this gap but face technical barriers: limited numbers of HDCOs cannot meet multi-well-plate demands, and long-term tracking accuracy is compromised by organoid overlapping/displacement during culture. Results: A multifunctional microfluidic chip (MOCiB-Chip) was developed, featuring a three-layer structure (fluidic layer, porous membrane, capture layer) and two independent regions for multi-concentration experiments. It integrates single-HDCO immobilization, culture, in situ observation, and on-chip drug mixing. Combined with an independently developed analysis program, Ciliary Beating Frequency Analysis Software (CBFAS), and a fluid control module, the automatic quantifying system for ciliary beating (AuCilia) was established. It achieved 7-day monitoring of human bronchial HDCOs and evaluated roflumilast at 0–100 nM: roflumilast at 10 nM increased beating frequency by 81.5 %, 1 nM by 27.8 %, while 100 nM caused cessation by day 2. cAMP measurements confirmed roflumilast's mechanism, validating the system. Significance: This strategy eliminates HDCO loss via on-chip integration of all procedures, resolving the conflict between limited samples and multi-condition tests. It improves tracking accuracy by avoiding organoid displacement/overlapping, overcoming multi-well-plate limitations. The system's low cost and high reproducibility accelerate drug screening (e.g., roflumilast) and enable precise study of cilia physiology, laying a key technical foundation for cilia-related disease research and clinical translation.

AB - Background: The beating of motile cilia plays an important role in many physiological processes. In recent years, the precise monitoring of ciliary beating has provided valuable insights into cilia-related diseases and their therapies. However, a gap persists between homogeneous in vitro cell models and heterogeneous in vivo ciliary systems. Human-derived ciliary organoids (HDCOs) show promise in bridging this gap but face technical barriers: limited numbers of HDCOs cannot meet multi-well-plate demands, and long-term tracking accuracy is compromised by organoid overlapping/displacement during culture. Results: A multifunctional microfluidic chip (MOCiB-Chip) was developed, featuring a three-layer structure (fluidic layer, porous membrane, capture layer) and two independent regions for multi-concentration experiments. It integrates single-HDCO immobilization, culture, in situ observation, and on-chip drug mixing. Combined with an independently developed analysis program, Ciliary Beating Frequency Analysis Software (CBFAS), and a fluid control module, the automatic quantifying system for ciliary beating (AuCilia) was established. It achieved 7-day monitoring of human bronchial HDCOs and evaluated roflumilast at 0–100 nM: roflumilast at 10 nM increased beating frequency by 81.5 %, 1 nM by 27.8 %, while 100 nM caused cessation by day 2. cAMP measurements confirmed roflumilast's mechanism, validating the system. Significance: This strategy eliminates HDCO loss via on-chip integration of all procedures, resolving the conflict between limited samples and multi-condition tests. It improves tracking accuracy by avoiding organoid displacement/overlapping, overcoming multi-well-plate limitations. The system's low cost and high reproducibility accelerate drug screening (e.g., roflumilast) and enable precise study of cilia physiology, laying a key technical foundation for cilia-related disease research and clinical translation.

KW - Ciliary beating

KW - Drug evaluation

KW - Human-derived organoid

KW - Microfluidic chip

UR - https://www.scopus.com/pages/publications/105023823482

U2 - 10.1016/j.aca.2025.344967

DO - 10.1016/j.aca.2025.344967

M3 - Article

AN - SCOPUS:105023823482

SN - 0003-2670

VL - 1384

JO - Analytica Chimica Acta

JF - Analytica Chimica Acta

M1 - 344967

ER -