Tissue scaffolds for electrically excitable cells

a tissue scaffold and excitable cell technology, applied in the field of tissue scaffolds, can solve the problems of limited predictive power of human clinical trials and post-market surveillance of preclinical data acquired through the use of rodent brain slices, high animal care and labor-intensive use costs, etc., and achieves improved predictive power, reduced cost, and the effect of improving the quality of li

Inactive Publication Date: 2016-09-01
INNOVATIVE SURFACE TECHNOLOGIES LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0029]In some aspects, inventive methods and articles can provide one or more advantages, such as an in vivo-like electrophysiology, superior predictive power, lower cost and higher throughput than acute slices, more reproducible results than acute slices, reproduction of circuits that

Problems solved by technology

However, the low number of brain slices that can be produced per animal, coupled with the high costs of animal care and their labor-intensive use, is a significant drawback to this approach.
Moreover, due to interspecies genetic variation, preclinical data acquired through the use of rodent brain slices will be of limited predictive power for human clinical trials and postmarket surveillan

Method used

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  • Tissue scaffolds for electrically excitable cells
  • Tissue scaffolds for electrically excitable cells
  • Tissue scaffolds for electrically excitable cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

Electrospinning of Random-Nonwoven and Aligned Photoreactive Nanofibers

[0156]Photoreactive polycaprolactone nanofibers were prepared by electrospinning solutions containing 10% poly(ε-caprolactone) (PCL, with an average molecular weight of 80 kDa, purchased from Sigma-Aldrich) and 0.1% Photoreactive Crosslinker in a 1:1 solution of tetrahydrofuran:N,N-dimethylformamide. Electrospinning procedure was as described in Example 2 of U.S. Patent Application Publication No. US 2014 / 0294783 A1 (Jie Wen et al., published 2 Oct. 2014). Random, nonwoven nanofiber mats (meshes) were collected as described in the referenced Example.

[0157]The formed nanofiber mats were removed, placed in a vacuum chamber for at least 48 hours to remove organic solvent residue, then stored in a desiccator. Fiber diameter, morphology and pore size of the dried nanofibers were characterized using light and scanning electron microscopy (SEM). FIG. 3A illustrates an SEM image of PCL nanofibers containing 1% Photoreact...

example 2

Photopatterning of Nanofiber Mats

[0160]Nanofiber mats were treated to create a surface that included a passivating polymer (PEG) surrounding discrete domains for cell culture and growth. Silver halide photolithographic masks were used to create the discrete cell culture domains.

[0161]Work was performed using sterile procedures. Materials were sterilized with 70% ethanol and illuminated with UV light (306 nm) for 3 minutes per side. Prior to photopatterning, photoreactive nanofiber mats were incubated in a solution of polyethylene glycol (PEG, MW 10K, Sigma) for 2 hours, and dried in the dark.

[0162]Complimentary silver halide photolithographic films (CAD / Art Services, Inc., Bandon, Oreg.) were used to conjugate polyethylene glycol (PEG) outside of the desired cell growth and attachment areas (see FIG. 5 and FIG. 1B). The mask sets were provided in oval shapes. Complementary masks enabled simultaneous light exposure to both sides of the nanofiber mat and ensure mask alignment and opti...

example 3

Cytocompatibility of Scaffolding Material

[0168]To investigate the ability of prepared scaffolding material to support cell adhesion and proliferation, samples were sterilized with 70% ethanol for 24 hours, washed extensively with PBS (0.1M, pH 7.4) and exposed to UV light for 40 minutes (CL 1000 ultraviolet crosslinker, UVP). Scaffold materials were seeded with myoblasts (C2C12 cells, ATCC), endothelial cells (BAEC (Bovine aortic endothelial cells), Lonza Biosciences), and neural cells to observe the ability of the nanofibrous scaffolds to support growth of these cells.

[0169]Muscle myoblast cells. FIG. 3B shows attachment and growth of C2C12 cells on nanofibrous scaffolds. The scaffolds were prepared as described in Example 1. An amount of the formed nanofibrous mesh was immersed in 20 ml of 50 mg / ml PAA aqueous solution for 30 minutes in a quartz round dish (Quartz Scientific, Inc., Fairport Harbor, Ohio). Mild agitation was applied to remove the air bubbles trapped in the nanofibe...

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Abstract

Inventive concepts relate generally to nanofibrous scaffolds useful for electrophysiological assays. Scaffolds include polymeric nanofibrous components and electrically excitable cells immobilized at a distinct cell seeding domains on the scaffold. Methods and kits including the scaffolds are also described.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application claims benefit under 35 U.S.C. §119(e) to U.S. Patent Application Ser. No. 61 / 890,193, entitled “CELL GROWTH SUBSTRATES CONTAINING OPEN FIELD ARCHITECTURES,” filed Oct. 12, 2013, the contents of which are incorporated herein in their entirety for all purposes.FIELD OF THE INVENTION[0002]Inventive concepts relate generally to the field of drug discovery and drug development for the screening of disease treatment.BACKGROUND[0003]The global drug discovery technologies market is projected to reach $19.9 Billion by 2017. In the primary methodology, High Throughput drug Screening process (HTS), massive libraries of chemical entities are evaluated for activity towards a target of interest, often expressed in a suitable cell line.[0004]This identification process is then followed by hit confirmation, hit expansion, and lead optimization for drug candidates. Additionally, high-content screening utilizing fluorescence, morphologica...

Claims

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Application Information

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IPC IPC(8): C12N11/08C12M1/34C12N5/00C12M1/12
CPCC12N11/08C12N5/0068C12M41/46C12M25/14C12N2533/30C12N11/089C12N11/096
Inventor GUIRE, ERICBAHR, CHRISTOPHER
Owner INNOVATIVE SURFACE TECHNOLOGIES LLC
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