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Microfabricated devices and high throughput assays for modulators of cell behavior

A technology for micro-manufacturing and regulators, which can be used in cell culture active agents, enzymology/microbiology devices, embryonic cells, etc., and can solve problems such as low throughput

Pending Publication Date: 2021-07-13
NOVOHEART LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] While the high-content nature of TS platforms shows promise for use in drug discovery and drug screening applications, most TS platforms are limited by their lower throughput compared to simpler platforms

Method used

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  • Microfabricated devices and high throughput assays for modulators of cell behavior
  • Microfabricated devices and high throughput assays for modulators of cell behavior
  • Microfabricated devices and high throughput assays for modulators of cell behavior

Examples

Experimental program
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Effect test

example 1

[0049] method

[0050] Preparation of cardiomyocytes

[0051] The hES2 cell line was used to differentiate human embryonic stem cells (hESCs) into human ventricular-like cardiomyocytes (hvCM) according to Novoheart's proprietary protocol based on the embryoid body approach [8]. Briefly, at 37 °C using 5% CO 2 Cells were maintained on matrigel-coated plates with mTeSR1. On the first day of differentiation, cells were digested to form a suspension in ultra-low attachment plates containing matrigel, 1 ng / ml bone morphogenic protein 4 (BMP4), and 10 μM ROCK inhibitor Y-27632 ( RI) small cell clusters persisting for 24 hours in mTeSR1. The medium was then replaced with StemPro-34 medium with GlutaMAX supplemented with 50 μg / mL ascorbic acid, 10 ng / mL Activin A, 10 ng / mL BMP4 and 5 μM RI. After 3 days, the cells were cultured for 4 days in StemPro-34 medium supplemented with 50 μg / mL ascorbic acid and 5 μM IWR-1. Thereafter, cell clusters were maintained in RPMI 1640 suppleme...

example 2

[0064] Force Measurement of Tissue on a Bending Column

[0065] Use bright-field microscopy to image the movement of the column over time due to the beating of the attached tissue strip ( image 3 A). Use custom LabVIEW code to track and record bar positions ( Figure 4 ). The position data was analyzed using a custom MATLAB code utilizing the beam bending equation to estimate the applied force ( Figure 4 )[9]. The beam bending equation is where F is the force, E is the Young's modulus of the column material, R is the radius, a is the height at which the force is applied on the column, L is the full column height, and δ is the displacement of the center of the column. It is estimated that the tissue is anchored at the center of the curved portion of the post. The system was engineered and aimed at centering tissue attachment at the center of the curvature and then visually confirming it afterward using biopsy cultures. Perform finite element modeling to confirm whet...

example 3

[0067] Fabrication and Assembly of Vertical Stacked Cardiac Tissue Culture Plates

[0068] To fabricate planar cardiac tissue prototype plates, a stacking or layering approach ( Figure 6 ), the method consisted of 1) bonding a thin silicone sheet to a commercial bottomless polystyrene cell culture plate, 2) laser cutting the thin silicone sheet in a desired pattern, 3) fabricating a tissue-forming trough layer, and 4) adhering the trough layer to the bottomless plate.

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Abstract

The disclosure provides a device for developing organized tissue strips, such as cardiac tissue strips, for high-throughput assays of functional performance and the methods involved in fabricating, assembling, implementing, utilizing and analyzing data from such assays. The disclosure further provides systems for constructing such devices, systems comprising those devices comprising cells and extracellular matrix material for developing organized tissue strips or comprising the devices and organized tissue strips. The disclosure further provides methods for assaying a property of a tissue strip, such as contractile force.

Description

technical field [0001] The present disclosure relates to the field of microfabrication and throughput methods for screening modulators of cellular behavior. Background technique [0002] Cardiotoxicity remains a key challenge in the development of new drugs, leading to significant drug attrition during phase I-III clinical trials and after marketing approval [1]. This failure is partly attributable to the lack of effective preclinical drug screening platforms. The current standard of preclinical screening utilizes animal models that have been shown to be insufficiently predictive of cardiotoxicity compared to humans due to interspecies differences in cardiac physiology [2]. To this end, researchers have focused considerable effort on the use of human pluripotent stem cell (hPSC)-derived cardiomyocytes (CMs) comprising induced pluripotent stem cells (iPSCs) reprogrammed from differentiated A preclinical drug screening tool for the physiological response of cardioactive comp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12M1/36
CPCC12M35/02C12M35/04C12M41/46G01N33/4833C12N5/0657B01L3/5085B01L2300/0663B01L2300/123B01L2300/163B01L2200/12G01N33/5014G01N33/5061G01N33/5073C12N2501/155C12N2501/727C12N2500/38C12N2501/15C12N2501/415C12N2506/02C12N2513/00C12N2533/90C12N2533/54C12N2502/09C12N2502/1329C12N5/0697C12N2500/02A61L27/3633A61L27/3834A61L27/3891C12M21/08
Inventor D·D·特兰李国玺Y·黑川E·G·罗伯茨K·D·科斯塔
Owner NOVOHEART LTD
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