Biocompatible, microscopy-compatible, resealable biopsy holder for microfluidic long-term cultivation and (chemo)therapy testing

Abstract

Biopsy holder (1) for receiving at least one tissue sample (2) under microfluidic conditions, comprising: ◯ a two-part housing (3, 4) which forms a receiving chamber (5) for the tissue sample; ◯ at least one inlet (6) and one outlet (7) for carrying out a continuous or intermittent perfusion flow of a culture medium through the receiving chamber; ◯ a sealing device (8) in the form of an integrated silicone O-ring, X-ring or sealing tape; ◯ The biopsy holder is made of biocompatible, sterilizable material and is manufactured using high-resolution 3D printing.

Description

[0001] Technical field: The invention relates to a microfluidic biopsy holder for long-term cultivation and microscopic analysis of tissue samples, in particular tumor biopsies, under dynamic perfusion conditions. The holder is suitable for use in incubators and microscopes and enables patient-specific experiments in modular Organon Chip systems.

[0002] State of the art: Conventional biopsy holders are usually statically constructed, difficult to access, and often incompatible with long-term experiments, continuous perfusion, and high-resolution microscopy. Solutions that combine standardized external dimensions (e.g., slide format), individually adaptable geometries for biopsies, and controlled, leak-free media flow are lacking. Furthermore, many existing systems are not resealable, lack long-term leak-proof performance, or are not suitable for modular use in automated analysis platforms.

[0003] US 2014 / 0113838 A1 discloses a microfluidic device used to predict drug effects on tumor biopsies. However, this system focuses on rapid test runs under simpler conditions and does not feature a resealable or individually configurable biopsy collection unit with flexible media routing.

[0004] WO 2021 / 058782 A1 describes a sample processing system with a microfluidic cartridge for the automated analysis of tissue samples. However, the device is based on closed, disposable components with limited microscopy compatibility and does not allow adaptation to different biopsy geometries or modular coupling of multiple tissue units.

[0005] Furthermore, US 10,087,422 B2 discloses so-called "organ-on-chip" systems that simulate the function of individual organs in vitro. However, these systems are primarily designed for synthetically produced cell cultures and not for the direct acquisition and long-term perfusion of real biopsies with individual geometries.

[0006] US Pat. No. 11,022,603 ​​B2 presents a microfluidic system for immunological drug assessment. While this system allows for a certain degree of interaction detection, it lacks a resealable structure for repeated sample access, modular coupling capability, and a specific adaptation for optical high resolution via an integrated coverslip.

[0007] US 9,149,806 B2 concerns microfluidic devices for cell separation and, in terms of its technical objective and design, is not comparable to a holder for long-term cultivation and microscopic observation of biopsy-based tissue.

[0008] The present invention goes beyond these known solutions by providing a flexibly configurable, biocompatible, and sterile, reusable system specifically tailored to the requirements of long-term cultivation of real patient samples under microfluidic conditions and their continuous microscopic analysis. Furthermore, the system allows the combination of multiple tissue units—such as tumor, liver, and kidney samples—in a connected modular setup for investigating patient-specific interactions and metabolic effects. Purpose of the invention:

[0009] The biopsy holder according to the invention enables the long-term cultivation and examination of tissue samples – especially tumor biopsies – under microfluidic conditions. The goal is to test the effects of different medications directly on patient material in order to make an individualized, optimized treatment decision. The invention allows for controlled observation of cell reactions in real time – both on target cells (e.g., tumor cells) and on healthy cell types from the same patient – ​​and thus supports the early assessment of therapeutic effects and potential side effects. Extended embodiments • The biopsy can be replaced by an organ equivalent generated from the patient's own cells, e.g., liver organoids, kidney organoids, or other cell cultures. These are supported by the geometry of the recording chamber and the customized manufacturing technology. • Multiple holders can be fluidically and mechanically connected in a separate modular system, e.g. using standardized coupling elements, to represent complex metabolic and systemic interactions, including the metabolism of drugs in an organ module (e.g. liver) and the effect of the metabolites on target tissue (e.g. tumor). • The modules can be configured to operate in a common microfluidic circuit or to be individually connected (parallel or serial configuration). • This can create a “life chip” system that simultaneously maps various aspects of individual physiology (e.g. liver, kidney and tumor response). Technical benefits • The combination of individual geometry adaptation, microfluidic control and modular connectivity can create in vitro conditions that enable investigations into physiologically realistic reactions of the patient material to therapeutic approaches. • Optical accessibility through integrated glass bottom or coatable, transparent surfaces allows continuous, high-resolution observation of cellular reactions - especially by fluorescence or confocal microscopy. • The reusable, hermetically sealed design allows long-term testing (214 days) under incubator conditions without media leakage or risk of contamination. • Enables controlled microfluidic experiments with high reproducibility. • Promotes personalized medicine through standardized integration into automated analysis processes. List of reference symbols 1 biopsy holder 2 tissue sample 3 Lower part 4 Top 5 Recording chamber for the tissue sample 6 Entrance 7 Outlet 8 Sealing device QUOTES CONTAINED IN THE DESCRIPTION

[0000] This list of documents submitted by the applicant was generated automatically and is included solely for the convenience of the reader. This list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions. Cited patent literature

[0000] US 2014 / 0113838 A1

[0003] WO 2021 / 058782 A1

[0004] US 10,087,422 B2

[0005] US 11,022,603 ​​B2

[0006] US 9,149,806 B2

[0007]

Claims

[1] Biopsy holder (1) for receiving at least one tissue sample (2) under microfluidic conditions, comprising: ◯ a two-part housing (3, 4) which forms a receiving chamber (5) for the tissue sample; ◯ at least one inlet (6) and one outlet (7) for carrying out a continuous or intermittent perfusion flow of a culture medium through the receiving chamber; ◯ a sealing device (8) in the form of an integrated silicone O-ring, X-ring or sealing tape; ◯ The biopsy holder is made of biocompatible, sterilizable material and is manufactured using high-resolution 3D printing. [2] Biopsy holder according to claim 1, characterized by that the sealing device is additionally reinforced by a transparent, adhesive sealing film element. [3] Biopsy holder according to one of the preceding claims, characterized bythat the recording chamber is individually adapted to the biopsy geometry. [4] Biopsy holder according to one of the preceding claims, characterized by that the housing has the external dimensions of a microscope slide. [5] Biopsy holder according to one of the preceding claims, characterized by that it contains an integrated transparent base or a fixed glass plate for high-resolution microscopy. [6] Biopsy holder according to one of the preceding claims, characterized by that there are several recording chambers, each with separate microfluidic circuits. [7] Biopsy holder according to one of the preceding claims, characterized by that the recording chambers are microfluidically connected to each other. [8] Biopsy holder according to one of the preceding claims, characterized by that biopsies can be replaced by patient-specific cultured organoids. [9] Biopsy holder according to one of the preceding claims, characterized by that several holders can be connected mechanically and fluidically in a separate module.

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