Hanging-drop three-dimensional cell culture device and methods employed therewith

The hanging-drop cell culture device addresses evaporation and scalability issues by providing stable droplet formation and direct imaging, enabling efficient 3D tissue generation and histology-compatible workflows.

WO2026069383A1 Publication Date: 2026-04-02JAGIRDAR RAJESH M
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Patent Information

Application Number
PCT/IN2025/051578
Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
Priority Date
2024-09-27
Filing Date
2025-09-27
Publication Date
2026-04-02

AI Technical Summary

Technical Problem

Existing hanging-drop cell culture methods face issues with evaporation, mechanical shock, sample intermixing, low throughput, and incompatibility with histology embedding, leading to scalability constraints and cumbersome analytical workflows.

Method used

A compact, microscope-compatible hanging-drop cell culture device with concave bottom regions and underside imaging cavities, enabling stable droplet formation, direct imaging, on-slide washing, and retrieval without centrifugation, and compatibility with histology workflows, using modular and scalable arrays.

Benefits of technology

Facilitates scalable generation and maintenance of 3D tissues with reduced evaporation, stable handling, and compatibility with histology embedding, supporting high-magnification observation and simplified retrieval processes.

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Abstract

A slide-like hanging-drop cell culture device (100) is disclosed for generating, maintaining, observing, and processing three-dimensional tissues. The device provides an array of culture cavities (102) with concave bottoms (103) configured to promote aggregation and, after re-inversion, to seat formed tissues for repeatable positioning. An underside imaging cavity (104) with an inner step (105) accommodates an immersion medium (162) to enable high-magnification observation from below, while the upper face supports coverslip-based imaging using a coverslip (160), thereby permitting bidirectional microscopy without transfer. A detachable cover (120) forms a closed arrangement and incorporates hydration features (124, 126) to reduce droplet evaporation during incubation. Optional accessories include a wash array (140) arranged to deliver fluids via conduits (142) and a wash-filter array (150) with conduits (152), extensions (154) and an internal filter (155) to retain fragments during processing. Methods of use include dispensing cell suspensions, forming hanging-drops, incubating, re-inverting for positioning, washing or permeabilizing on the slide, imaging from either side, and retrieving (182) or embedding (170) tissues for downstream analysis with optional lift- out using forceps (180).
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