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Macro tissue explant, methods and uses therefor

a micro-tissue and explant technology, applied in the field of micro-tissue explant, methods, can solve the problems of difficult study of mammalian organs, inability to fully recapture the architecture and function of a tissue, and general incompatibility of cultures with high-throughput systems

Pending Publication Date: 2019-02-28
THE BRIGHAM & WOMEN S HOSPITAL INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent describes a substrate that has a large number of small wells covered by tissue. These wells can be 6, 12, 24, 48, 96, 384, or 1536 micrometers in size. The substrate does not contain any materials that are not natural to the tissue. The tissue also has a minimal amount of blood content. The technical effect of this invention is a structured biological substrate that can provide a more accurate and reliable environment for the growth and expansion of cells.

Problems solved by technology

Mammalian organs are challenging to study as they are fairly inaccessible to experimental manipulation and optical observation.
However, these systems fail to fully recapitulate the architecture and functionality of a tissue.
Furthermore, these cultures are generally incompatible with high-throughput systems.

Method used

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  • Macro tissue explant, methods and uses therefor
  • Macro tissue explant, methods and uses therefor
  • Macro tissue explant, methods and uses therefor

Examples

Experimental program
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example 1

ultivation System for Intestinal Tissue

[0528]To determine whether intestinal tissue could be cultivated in a manner that maintained viability and in vivo architecture, small intestinal tissue was isolated from freshly procured intact gastrointestinal tract from pigs. The viability of intestinal tissue explants was found to be dependent on specific media compositions (FIG. 1A). No difference in viability was observed between tissue cultured in DMEM F12 alone and DMEM F12 with FBS or EGF added. In addition, FIG. 1B shows pictures of the luminal side of the small intestinal tissue explants with or without media cultivation after 7 days, indicating the explants survived best with media. The stroma was found to be essential in maintaining the cell survival of the intestinal epithelium in ex vivo cultivation (FIG. 1C). Confocal analysis of sectioned intestinal tissue explants that were stained with Dapi (blue, cell nucleus), Phalloidin (green, F-actin, Wheat Germ Agglutinin (Plasma membra...

example 2

l Tissue Explant Platform Development

[0537]Next, the use of the intestinal tissue explant from Example 1 in a high-throughput platform was investigated. An interface platform for intestinal tissue explant cultivation that enabled high-throughput intestinal drug perfusion measurements combined with long-term tissue cultivation capability was designed. Specifically, a broad range of different designs and materials for potential interfacing systems was systematically assessed. As shown in FIG. 2A, a system that enables low sample variability, tissue viability maintenance, rapid assembly and compatibility with robotic handling was developed. The design consisted of an upper device that compartmentalized the intestinal tissue in a 96 multi-well plate format. The tissue formed the bottom of the multi-well plate and was sealed off around each of the 96 wells by using an additional device underneath the tissue. The system was enclosed by a case that enabled adjustable pressure to maintain t...

example 3

l Tissue Explant System Intestinal Absorption Validation

[0546]The Food and Drug Administration (FDA), recommends using drugs approved for oral administration with human clinical pharmacokinetic data to validate the in vivo predictability of an in vitro intestinal perfusion system (Waiver of In Vivo Bioavailability and Bioequivalence Studies for Immediate-Release Sold Oral Dosage Forms Based on a Biopharmaceutics Classification System, U.S. Dep. Heal. Hum. Serv. Food drug Adm. Cent. Drug Eval. Res., 2000). Therefore, to confirm the use of the intestinal tissue explant as a system for predicting intestinal absorption, the perfusion of 60 model drugs was analyzed. Specifically, drugs from the 4 Biopharmaceutical Classification System (BCS) classes (16 BCs class I, 13 BCS class II, 15 BCS class III, and 12 BCS class IV), along with 4 dextran-based control substances, were used. The intestinal perfusion data obtained from the intestinal tissue explant system was compared to human intesti...

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Abstract

Tissue explants of the gastrointestinal tract are provided. Methods of making and using the tissue explants are also provided, along with substrates designed for the tissue explants described.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Provisional Application Ser. No. 62 / 476,181, filed on Mar. 24, 2017; and U.S. Provisional Application Ser. No. 62 / 560,485, filed on Sep. 19, 2017. The entire contents of the above-referenced applications are incorporated herein by this reference.BACKGROUND[0002]Mammalian organs are challenging to study as they are fairly inaccessible to experimental manipulation and optical observation. Recent advances in three-dimensional (3D) culture techniques, coupled with the ability to independently manipulate genetic and microenvironmental factors, have enabled the real-time study of mammalian tissues.[0003]However, these systems fail to fully recapitulate the architecture and functionality of a tissue. Furthermore, these cultures are generally incompatible with high-throughput systems. Therefore, a need remains for systems that can mimic the in vivo architecture and function of a tissue and be subjected to high-throughput ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/50C12N5/00C12N5/071
CPCG01N33/5088C12N5/0062C12N5/0679C12N2513/00C12N2503/02C12N2503/04C12N2510/00C12N2531/00
Inventor TRAVERSO, CARLO GIOVANNIVON ERLACH, THOMAS CHRISTIANLANGER, ROBERT S.SAXTON, SARAHMINAHAN, DANIEL
Owner THE BRIGHAM & WOMEN S HOSPITAL INC
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