Vitro characterization of cell injury due to mechanical blunt
Inventors
Raphael, Marc P. • Kang, Wonmo
Assignees
Publication Number
US-11702622-B2
Publication Date
2023-07-18
Expiration Date
2040-08-21
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Abstract
A system and method for studying cell injury mechanisms by applying biologically relevant mechanical impact to in vitro cell culture are disclosed. This approach is for maintaining consistent in vitro conditions during experiments, accommodating multiple cell populations, and monitoring each in real-time while achieving amplitude and time scale of input acceleration that mimic blunt injury cases. These multiplexed, environmental control capabilities enable characterizing the relationships between mechanical impact and cell injury in multivariate biological systems.
Core Innovation
The invention discloses a system and method for in vitro characterization of cell injury due to mechanical blunt, employing a well-controlled mechanical impact to live cells cultured in an in vitro setup compatible with live cell microscopy. This approach enables application of well-characterized and reproducible acceleration-induced pressure gradients, while maintaining consistent in vitro conditions over long-term and environmentally controlled observations. It facilitates multiplexed monitoring of multiple cell populations to investigate cell viability, mitosis, and injury mechanisms such as membrane permeabilization caused by mechanical impact.
The problem addressed is the limited understanding of post-trauma cellular damage mechanisms due to a lack of standard testing procedures and complexity in interpreting animal models. Existing in vitro platforms face technical challenges in applying biologically relevant mechanical impacts with controlled forces and temporal profiles. Moreover, conventional wound healing assays fail to mimic dynamic blunt injury processes accurately, lacking multiplexing and long-term observation capabilities. The invention seeks to overcome these limitations by providing a multiplexed, consistent, and quantitative in vitro platform for studying blunt injury at cellular and molecular levels.
Claims Coverage
The independent claims define two main inventive features covering both a method and a system for in vitro characterization of cell injury due to mechanical blunt.
In vitro method for characterizing cell injury due to mechanical blunt
The method involves imaging live cell populations in a cell culture setup within an incubator using live cell imaging instruments before and after applying a controlled mechanical impact. The setup includes a holder incorporating a first foam above and a second foam below the cell culture. During impact, measurement of velocity, acceleration, impact force, and cavitation-induced pressure is performed concurrently with optical monitoring. The dynamic cellular response is monitored for up to two weeks, facilitating mapping of cell injury relationships over time.
System for in vitro characterization of cell injury due to mechanical blunt
The system comprises an impact system including a holder with a first foam above and a second foam below, at least one camera, an incubator, a live cell imaging instrument, and a cell culture setup containing live cell populations. The cell culture setup is maintained in the incubator for imaging before and after impact and mounted in the holder during impact. The system also includes a data acquisition system for capturing impact-related parameters.
The claims cover an integrated method and system capable of applying biologically relevant mechanical impacts with precise control and measurement, enabling multiplexed, long-term live cell imaging to study blunt injury mechanisms in vitro.
Stated Advantages
Capability to apply well-characterized and reproducible acceleration-induced pressure gradients compatible with long-term and environmentally controlled live-cell microscopy.
Permits close monitoring of multiple cell populations, down to individual cells, to deduce effects on cell viability, mitosis, and potential molecular injury mechanisms such as membrane permeabilization.
Enables continuous optical observation of multiple cell cultures simultaneously without removing them from incubators, reducing batch-to-batch and population heterogeneity effects.
Supports high throughput analyses and multiplexed studies improving statistical significance for heterogeneous cell responses.
Documented Applications
Studying cellular damage mechanisms related to blunt liver trauma and traumatic brain injuries by applying biologically relevant mechanical impacts to in vitro cell cultures.
Investigating the role of acceleration and cavitation-induced pressure in trauma-related cell injury in multivariate biological systems.
Enabling live cell microscopy-based monitoring of cell viability, mitosis, and membrane integrity before and after mechanical impact.
Quantitative assessment of injury criteria related to mechanical blunt impacts including acceleration and cavitation dynamics using multiplexed in vitro platforms.
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