In-vitro contractile force indicator
Inventors
Grasman, Jonathan • Pridmore, Jessica • Martin, Cassandra • Coombs, Katherine • Sicherer, Sydnee
Assignees
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Publication Number
US-12385898-B2
Publication Date
2025-08-12
Expiration Date
Abstract
Devices and methods to measure cells and/or tissue's contractile force are disclosed. Included is a mount with rigid, and non-rigid posts sized to flex. Determined is force exerted by contractile cells and tissues in a multiwell plate. The device is designed to fit inside individual wells with posts directed downwards. Posts are attached to a 3D printed circular mount with tabs for depth within the well. The mount has a window for medium changes while the device is positioned inside the well. The cells are seeded within a hydrogel. As the hydrogel condenses, cells/tissue wrap around the post's outside pulling non-rigid post toward rigid post. Inverted light microscope is used to determine deflection of non-rigid post inside the multiwell plate. Movement of the non-rigid post is measured using an acrylic ruler on an underside of the multiwell plate. Contractile forces of cells/tissue are determined using cantilever mechanics.
Core Innovation
The invention provides an in-vitro contractile force indicator for a growing specimen in a culture plate well. The indicator includes a first mount having a cylindrical shape to fit inside a well of a culture plate, and the growing specimen is configured to grow and wrap around a first rigid post and a first non-rigid post positioned vertically and pointing downward into the well.
The first rigid post has no deflection or no deformation when an external force of at least 30 μN is applied by the growing specimen, remains stationary with respect to the first mount, and serves as an anchor when the first non-rigid post deflects. The first non-rigid post is interchangeably attached to the first mount, parallel to the first rigid post, and has a deflection or a deformation when the external force of at least 30 μN is applied.
The deflection of the non-rigid post is used for calculating a force of contraction of the specimen. The document describes non-destructive, real-time measurement in connection with cells and/or tissues, including a hydrogel specimen that contracts and pulls the non-rigid post toward the rigid post, and the indicator is supported by cantilever mechanics for force calculation and calibration based on mechanical behavior and elastic modulus constraints.
Claims Coverage
The independent claim coverage includes the cylindrical mount, the stationary rigid anchor post, the parallel downward-pointing deflecting non-rigid post, and deflection-based calculation of specimen contraction force. Additional claimed features include an under-plate measurement ruler, autoclavable materials for the posts, a quantified PDMS elastic modulus after autoclave, real-time contractile-force measurement with cellular specimens, and a drug discovery context using Duchenne muscular dystrophy patient-derived myoblasts.
Cylindrical culture-plate mount for a growing specimen
A first mount having a cylindrical shape to fit inside a well of a culture plate that contains a growing specimen.
Stationary vertical rigid anchor post with no deflection at ≥30 μN
A first rigid post integral to the first mount, positioned vertically and pointing downward into the well, remaining stationary with no deflection or no deformation when an external force of at least 30 μN is applied by the growing specimen.
Parallel downward-pointing non-rigid deflecting post interchangeably attached
A first non-rigid post interchangeably attached to the first mount, parallel to the first rigid post, pointing downward into the well, and having a deflection or a deformation when the external force of at least 30 μN is applied by the growing specimen.
Anchor-to-deflection mechanism for calculating contraction force
The first rigid post serves as an anchor when the first non-rigid post undergoes deflection due to specimen contraction wrapping around the rigid and non-rigid posts, wherein the deflection is for calculating a force of contraction of the specimen.
Under-plate ruler patterns for measuring post deflection
A ruler positioned underneath a multiwell culture plate, including a plurality of circular patterns and a parallel pattern of hatch marks, disposed under the multiwell culture plate to measure the deflection of the first non-rigid post.
Autoclavable rigid and non-rigid post materials
The first rigid post and the first non-rigid post are formed from autoclavable materials, with the rigid components including 2-hydroxyethyl methacrylate, phenyl bis(2,4,6-trimethyl benzoyl)-phosphine oxide, and urethane dimethacrylate, and the non-rigid post including polydimethylsiloxane (PDMS).
Quantified PDMS elastic modulus after autoclave
The PDMS has an elastic modulus of 1.20 ± 0.60 MPa after 1× autoclave.
Real-time contractile-force measurement driven by cell forces during growth
For a specimen containing cells, the real-time contractile-force indicator measures deflection caused by cell forces constricting the first rigid and the first non-rigid posts as the cells grow.
Drug discovery use with Duchenne muscular dystrophy patient-derived myoblasts
Measuring how different drugs or drug dosages affect muscular strength or recovery using a specimen containing cells or tissue of Duchenne muscular dystrophy patient-derived myoblasts that exert contractile forces.
Overall, the claims cover a contractile force indicator that converts contraction-induced deflection of a non-rigid post, anchored by a rigid stationary post, into a calculated force of contraction, with additional coverage for an under-plate measurement ruler, specified autoclavable materials, a quantified PDMS elastic modulus constraint after autoclave, real-time cellular measurement, and a drug-discovery context using Duchenne muscular dystrophy patient-derived myoblasts.
Stated Advantages
Non-destructive, real-time measurement of contractile force using deflection of a non-rigid post.
The deflection is used for calculating a force of contraction of the specimen.
Enables evaluation of how different drugs or drug dosages affect muscular strength or recovery in a Duchenne muscular dystrophy context.
Documented Applications
Drug discovery for muscular strength or recovery, including evaluating effects of different drugs or drug dosages using a specimen containing Duchenne muscular dystrophy patient-derived myoblasts.
Real-time contractile-force measurement for a specimen containing cells as the cells grow.
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