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Cell culture system

一种细胞培养、细胞的技术，应用在组织细胞/病毒的培养装置、胃肠细胞、动物细胞等方向，能够解决活微生物不能生长、未重现活肠等问题

Active Publication Date: 2014-01-08
PRESIDENT & FELLOWS OF HARVARD COLLEGE
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  • Abstract
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  • Claims
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Problems solved by technology

However, none of the existing in vitro intestinal models reproduce the mechanically active microenvironment (peristaltic motion and intraluminal fluid flow) of the living intestine, which is essential for normal organ physiology. 15 and the development of Crohn's disease and other bowel disorders 16-17 is very critical
Another limitation of existing in vitro intestinal models is that viable microorganisms have not yet been able to grow over extended periods of time on the luminal surface of cultured intestinal epithelium as is often the case in living intestines.

Method used

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Embodiment 1

[0203] The "gut-on-a-chip" can be a microfluidic system containing a monolayer of cultured human intestinal epithelial cells and a monolayer of human endothelial cells from capillaries and / or lymphatic lacteal ducts coated with an extracellular matrix (ECM ) separated by a flexible porous membrane (which can undergo rhythmic mechanical twisting similar to intestinal motility (e.g., peristalsis and segmental movement)), which was designed to reproduce the tissue-tissue interface and microstructure of the human intestine ( figure 1 ). The goal of the "Gut on a Chip" project is to provide a robust, reproducible and predictable in vitro platform for human responses to orally delivered compounds, therapeutics, nutraceuticals, functional foods, pathogens and toxins. The gut-on-a-chip should be useful in a wide range of applications in the following fields: pharmacology, toxicology, drug development, drug delivery, drug metabolism, drug-drug interactions, drug bioavailability, drug ...

Embodiment 2

[0226] The development of living cell-based in vitro intestinal models (mimicking the mechanisms, structure, absorption, transport, and pathophysiological properties of the human intestine and its important microbial symbionts) could expedite drug development and potentially replace animal experiments. This paper describes one embodiment of a biomimetic "human intestine-on-a-chip" microdevice consisting of two microfluidic channels composed of porous flexible membranes coated with extracellular matrix (ECM). The membrane separates and is lined with human intestinal epithelial (Caco-2) cells to mimic the complex structure and physiology of the living intestine. Reconstruct the intestinal microenvironment through the following steps: make low shear stress (0.02dyne / cm 2 ) fluid at a low flow rate (30 μL / hr) flowed through the microchannel, and a periodic strain (10%; 0.15 Hz) imitating physiological peristaltic motion was applied. Under these conditions, the columnar epithelium...

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Abstract

The embodiments of the invention described herein relate to systems and methods for culturing and / or maintaining intestinal cells, tissues and / or organoids in vitro. The cells, tissues and / or organoids cultured according to the methods and systems described herein can mimic or reproduce natural intestinal epithelial structures and behavior as well as support co-culture of intestinal microflora.

Description

[0001] Cross References to Related Applications [0002] This application claims priority under 35 U.S.C. 119(e) to U.S. Provisional Patent Application Serial No. 61 / 447,540 (filed February 28, 2011), the contents of which are each incorporated herein by reference in their entirety. [0003] governmental support [0004] This invention was made in part with US Government support under grant ES016665-01A1 from the National Institute of Environmental Health Sciences. The US Government has certain rights in this invention. technical field [0005] The systems and methods of the invention described herein relate to the in vitro culture and maintenance of intestinal organoids. Background technique [0006] Drug development hampered by reliance on use of animal models, which are costly, labor-intensive, time-consuming, and ethically questionable 1 . But a bigger concern is that animal models often fail to predict the results obtained in humans 2-3 , which are unique problems ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/02C12N5/071C12N1/20C12N1/14
CPCC12N1/14C12N1/20C12M25/02C12M21/08C12M35/04C12M35/08C12M23/08C12M23/38C12M27/02C12M29/04C12M29/06C12M41/12C12M41/26C12M23/20C12M23/26C12M23/34C12M41/40C12M41/44C12N5/0679C12N5/068C12N5/0696C12N5/0697
Inventor 唐纳德·E·英格贝尔金贤政
Owner PRESIDENT & FELLOWS OF HARVARD COLLEGE
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